Benzodiazepine derivatives, compositions, and methods for treating cognitive impairment

ABSTRACT

This invention relates to benzodiazepine derivatives, compositions comprising therapeutically effective amounts of those benzodiazepine derivatives and methods of using those derivatives or compositions in treating cognitive impairment associated with central nervous system (CNS) disorders. In particular, it relates to the use of α5-containing GABA A  receptor agonist (e.g., a α5-containing GABA A  receptor positive allosteric modulator) as described herein in treating cognitive impairment associated with central nervous system (CNS) disorders in a subject in need or at risk thereof, including, without limitation, subjects having or at risk for age-related cognitive impairment, Mild Cognitive Impairment (MCI), amnestic MCI (aMCI), Age-Associated Memory Impairment (AAMI), Age Related Cognitive Decline (ARCD), dementia, Alzheimer&#39;s Disease (AD), prodromal AD, post traumatic stress disorder (PTSD), schizophrenia, bipolar disorder, amyotrophic lateral sclerosis (ALS), cancer-therapy-related cognitive impairment, mental retardation, Parkinson&#39;s disease (PD), autism spectrum disorders, fragile X disorder, Rett syndrome, compulsive behavior, and substance addiction.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.17/028,917, filed Sep. 22, 2020, now U.S. Pat. No. 11,312,721 issuedApr. 26, 2022 which is a divisional application of U.S. patentapplication Ser. No. 15/736,697, filed Dec. 14, 2017, now U.S. Pat. No.10,815,242 issued Oct. 27, 2020 which is a national stage applicationunder 35 U.S.C. § 371 of International Application PCT/US2016/038224,filed Jun. 17, 2016 (expired), which claims priority from U.S.Provisional Patent Application 62/182,336, filed Jun. 19, 2015(expired). Each of the foregoing applications is incorporated herein byreference in their entireties.

STATEMENT OF GOVERNMENT SUPPORT

This invention was made with government support under Grant No. U01AG041140 awarded by the National Institutes of Health (NIH), and inparticular, its National Institute on Aging (NIA) division, an agency ofthe United States Government. The United States Government has certainrights in the invention.

FIELD OF THE INVENTION

The invention relates to compounds, compositions and methods fortreating cognitive impairment associated with central nervous system(CNS) disorders in a subject in need of treatment or at risk of saidcognitive impairment.

BACKGROUND OF THE INVENTION

Cognitive ability may decline as a normal consequence of aging or as aconsequence of a central nervous disorder.

For example, a significant population of elderly adults experiences adecline in cognitive ability that exceeds what is typical in normalaging. Such age-related loss of cognitive function is characterizedclinically by progressive loss of memory, cognition, reasoning, andjudgment. Mild Cognitive Impairment (MCI), Age-Associated MemoryImpairment (AAMI), Age-Related Cognitive Decline (ARCD) or similarclinical groupings are among those related to such age-related loss ofcognitive function. According to some estimates, there are more than 16million people with AAMI in the U.S. alone (Barker et al., 1995), andMCI is estimated to affect 5.5-7 million in the U.S. over the age of 65(Plassman et al., 2008).

Cognitive impairment is also associated with other central nervoussystem (CNS) disorders, such as dementia, Alzheimer's Disease (AD),prodromal AD, post traumatic stress disorder (PTSD), schizophrenia,bipolar disorder (in particular, mania), amyotrophic lateral sclerosis(ALS), cancer-therapy-related cognitive impairment, mental retardation,Parkinson's disease (PD), autism spectrum disorders, fragile X disorder,Rett syndrome, compulsive behavior, and substance addiction.

There is, therefore, a need for effective treatment of cognitiveimpairment associated with central nervous system (CNS) disorders and toimprove cognitive function in patients diagnosed with, for example,age-related cognitive impairment, MCI, amnestic MCI, AAMI, ARCD,dementia, AD, prodromal AD, PTSD, schizophrenia or bipolar disorder (inparticular, mania), amyotrophic lateral sclerosis (ALS),cancer-therapy-related cognitive impairment, mental retardation,Parkinson's disease (PD), autism spectrum disorders, fragile X disorder,Rett syndrome, compulsive behavior, and substance addiction and similarcentral nervous system (CNS) disorders with cognitive impairment or atrisk of developing them.

GABA_(A) receptors (GABA_(A) R) are pentameric assemblies from a pool ofdifferent subunits (α1-6, β1-3, γ1-3, δ, ε, π, θ) that form a Cl−permeable channel that is gated by the neurotransmitter γ-aminobutyricacid (GABA). Various pharmacological effects, including anxietydisorders, epilepsy, insomnia, pre-anesthetic sedation, and musclerelaxation, are mediated by different GABA_(A) subtypes.

Various studies have demonstrated that reduced GABA signaling is linkedto various CNS disorders with cognitive impairment. In particular, theα5-containing GABA_(A) Rs, which are relatively sparse in the mammalianbrain, play a role in modifying learning and memory. Previous studiesdemonstrated a reduction of hippocampal expression of the α5 subunit ofthe GABA_(A) receptor in rats with age-related cognitive decline (seeInternational Patent Publication WO 2007/019312). Such results suggestthat upregulation of α5-containing GABA_(A) R function may be effectivein the treatment of cognitive impairment associated with said CNSdisorders.

Thus, there is a need for positive allosteric modulators ofα5-containing GABA_(A) R that are useful in therapeutic preparations forthe treatment of cognitive impairment associated with said CNSdisorders.

SUMMARY OF THE INVENTION

The present invention addresses the aforementioned need by providing acompound of formula I:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   U and the two carbon atoms designated by α and β together form a 5-    or 6-membered aromatic ring having 0-2 nitrogen atoms;-   A is C, CR⁶, or N;-   B and F are each independently selected from C, CR⁶, and N, wherein    B and F cannot both be N;-   D is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   E is N, NR⁷, CR⁶ or C(R⁶)₂;-   W is N, NR⁷, CR⁶ or C(R⁶)₂;-   X is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   Y and Z are each independently selected from C, CR⁶, and N, wherein    Y and Z cannot both be N;-   V is C or CR⁶,-   or when Z is C or CR⁶, V is C, CR⁶, or N;-   wherein when the ring formed by X, Y, Z, V and W is

then R² is —OR⁸, —SR⁸, —(CH₂)_(n)OR⁸, —(CH₂)_(n)O(CH₂)_(n)R⁸,—(CH₂)_(p)R⁸ and —(CH₂)_(n)N(R″)R¹⁰; and wherein R² is independentlysubstituted with 0-5 R′;

-   m and n are independently integers selected from 0-4;-   p is an integer selected from 2-4;-   each occurrence of the bond “    ” is either a single bond or a double bond; each occurrence of R¹,    R², R⁴, and R⁵ are each independently selected from:    -   halogen, —R, —OR, —NO₂, —NCS, —CN, —CF₃, —OCF₃, —SiR₃, —N(R)₂,        —SR, —SOR, —SO₂R, —SO₂N(R)₂, —SO₃R, —(CR₂)₁₋₃R, —(CR₂)₁₋₃—OR,        —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃R, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃OR, —C(O)R,        —C(O)C(O)R, —C(O)CH₂C(O)R, —C(S)R, —C(S)OR, —C(O)OR,        —C(O)C(O)OR, —C(O)C(O)N(R)₂, —OC(O)R, —C(O)N(R)₂, —OC(O)N(R)₂,        —C(S)N(R)₂, —(CR₂)₀₋₃NHC(O)R, —N(R)N(R)COR, —N(R)N(R)C(O)OR,        —N(R)N(R)CON(R)₂, —N(R)SO₂R, —N(R)SO₂N(R)₂, —N(R)C(O)OR,        —N(R)C(O)R, —N(R)C(S)R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂,        —N(COR)COR, —N(OR)R, —C(═NH) N(R)₂, —C(O)N(OR)R, —C(═NOR)R,        —OP(O)(OR)₂, —P(O)(R)₂, —P(O)(OR)₂, and —P(O)(H)(OR);-   R³ is absent or is selected from:    -   halogen, —R, —OR, —NO₂, —NCS, —CN, —CF₃, —OCF₃, —SiR₃, —N(R)₂,        —SR, —SOR, —SO₂R, —SO₂N(R)₂, —SO₃R, —(CR₂)₁₋₃R, —(CR₂)₁₋₃—OR,        —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃R, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃OR, —C(O)R,        —C(O)C(O)R, —C(O)CH₂C(O)R, —C(S)R, —C(S)OR, —C(O)OR,        —C(O)C(O)OR, —C(O)C(O)N(R)₂, —OC(O)R, —C(O)N(R)₂, —OC(O)N(R)₂,        —C(S)N(R)₂, —(CR₂)₀₋₃NHC(O)R, —N(R)N(R)COR, —N(R)N(R)C(O)OR,        —N(R)N(R)CON(R)₂, —N(R)SO₂R, —N(R)SO₂N(R)₂, —N(R)C(O)OR,        —N(R)C(O)R, —N(R)C(S)R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂,        —N(COR)COR, —N(OR)R, —C(═NH) N(R)₂, —C(O)N(OR)R, —C(═NOR)R,        —OP(O)(OR)₂, —P(O)(R)₂, —P(O)(OR)₂, and —P(O)(H)(OR);-   each R⁶ is independently —H or —(C1-C6)alkyl;-   each R⁷ is independently —H or —(C1-C6)alkyl;-   each R⁸ is independently —(C1-C6)alkyl, —(C3-C10)-cycloalkyl,    (C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each    occurrence of R⁸ is independently substituted with 0-5 R′;-   each R¹⁰ is independently —(C3-C10)-cycloalkyl, 3- to 10-membered    heterocyclyl-, (C6-C10)-aryl, or 5- to 10-membered heteroaryl,    wherein each occurrence of R¹⁰ is independently substituted with 0-5    R′;-   each R is independently selected from:    -   H—,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl-,    -   (C3-C10)-cycloalkenyl-,    -   [(C3-C10)-cycloalkyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkyl]-O—(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-O—(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C6-C10)-aryl-O—(C1-C12)aliphatic-,    -   (C6-C10)-aryl-N(R″)—(C1-C12)aliphatic-,    -   3- to 10-membered heterocyclyl-,    -   (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-N(R″)—(C1-C12)aliphatic-,    -   5- to 10-membered heteroaryl-,    -   (5- to 10-membered heteroaryl)-(C1-C12)-aliphatic-,    -   (5- to 10-membered heteroaryl)-O—(C1-C12)-aliphatic-; and    -   (5- to 10-membered heteroaryl)-N(R″)—(C1-C12)-aliphatic-;-   wherein said heterocyclyl has 1-4 heteroatoms independently selected    from N, NH, O, S, SO, and SO₂, and said heteroaryl has 1-4    heteroatoms independently selected from N, NH, O, and S;-   wherein each occurrence of R is independently substituted with 0-5    R′;-   or when two R groups bound to the same atom, the two R groups may be    taken together with the atom to which they are bound to form a 3- to    10-membered aromatic or non-aromatic ring having 0-4 heteroatoms    independently selected from N, NH, O, S, SO, and SO₂, wherein said    ring is optionally substituted with 0-5 R′, and wherein said ring is    optionally fused to a (C6-C10)aryl, 5- to 10-membered heteroaryl,    (C3-C10)cycloalkyl, or a 3- to 10-membered heterocyclyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;    wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl, 3- to    6-membered heterocyclyl, 5- to 10-membered heteroaryl-,    (C6-C10)-aryl-, (5- to 10-membered heteroaryl)-(C1-C6)-alkyl-,    (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to 10-membered    heteroaryl)-O—(C1-C6)-alkyl-, and (C6-C10)-aryl-O—(C1-C6)-alkyl-,    wherein each occurrence of R″ is independently substituted with 0-3    substituents selected from: halogen, —R^(∘), —OR^(∘), oxo,    —CH₂OR^(∘), —CH₂NR^(∘) ₂, —C(O)N(R^(∘))₂, —C(O)OR^(∘), —NO₂, —NCS,    —CN,    —CF₃, —OCF₃ and —N(R^(∘))₂, wherein each occurrence of R^(∘) is    independently selected from: —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl,    3- to 6-membered heterocyclyl, 5- to 10-membered heteroaryl-, and    (C6-C10)-aryl-.

Some embodiments of this application provide a compound of formula I:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   U and the two carbon atoms designated by α and β together form a 5-    or 6-membered aromatic ring having 0-2 nitrogen atoms;-   A is C, CR⁶, or N;-   B and F are each independently selected from C, CR⁶, and N, wherein    B and F cannot both be N;-   D is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   E is N, NR⁷, CR⁶ or C(R⁶)₂;-   W is N, NR⁷, CR⁶ or C(R⁶)₂;-   X is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   Y and Z are each independently selected from C, CR⁶, and N, wherein    Y and Z cannot both be N;-   V is C or CR⁶,-   or when Z is C or CR⁶, V is C, CR⁶, or N;-   wherein when the ring formed by X, Y, Z, V and W is

then R² is —OR⁸, —SR⁸, —(CH₂)_(n)OR⁸, —(CH₂)_(n)O(CH₂)_(n)R⁸,—(CH₂)_(p)R⁸ and —(CH₂)_(n)N(R″)R¹⁰; and wherein R² is independentlysubstituted with 0-5 R′;

-   m and n are independently integers selected from 0-4;-   p is an integer selected from 2-4;-   each occurrence of the bond “    ” is either a single bond or a double bond;-   each occurrence of R¹, R², R⁴, and R⁵ are each independently    selected from:    -   halogen, —R, —OR, —NO₂, —NCS, —CN, —CF₃, —OCF₃, —SiR₃, —N(R)₂,        —SR, —SOR, —SO₂R, —SO₂N(R)₂, —SO₃R, —(CR₂)₁₋₃R, —(CR₂)₁₋₃—OR,        —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃R, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃OR, —C(O)R,        —C(O)C(O)R, —C(O)CH₂C(O)R, —C(S)R, —C(S)OR, —C(O)OR,        —C(O)C(O)OR, —C(O)C(O)N(R)₂, —OC(O)R, —C(O)N(R)₂, —OC(O)N(R)₂,        —C(S)N(R)₂, —(CR₂)₀₋₃NHC(O)R, —N(R)N(R)COR, —N(R)N(R)C(O)OR,        —N(R)N(R)CON(R)₂, —N(R)SO₂R, —N(R)SO₂N(R)₂, —N(R)C(O)OR,        —N(R)C(O)R, —N(R)C(S)R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂,        —N(COR)COR, —N(OR)R, —C(═NH) N(R)₂, —C(O)N(OR)R, —C(═NOR)R,        —OP(O)(OR)₂, —P(O)(R)₂, —P(O)(OR)₂, and —P(O)(H)(OR);-   R³ is absent or is selected from:    -   halogen, —R, —OR, —NO₂, —NCS, —CN, —CF₃, —OCF₃, —SiR₃, —N(R)₂,        —SR, —SOR, —SO₂R, —SO₂N(R)₂, —SO₃R, —(CR₂)₁₋₃R, —(CR₂)₁₋₃—OR,        —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃R, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃OR, —C(O)R,        —C(O)C(O)R, —C(O)CH₂C(O)R, —C(S)R, —C(S)OR, —C(O)OR,        —C(O)C(O)OR, —C(O)C(O)N(R)₂, —OC(O)R, —C(O)N(R)₂, —OC(O)N(R)₂,        —C(S)N(R)₂, —(CR₂)₀₋₃NHC(O)R, —N(R)N(R)COR, —N(R)N(R)C(O)OR,        —N(R)N(R)CON(R)₂, —N(R)SO₂R, —N(R)SO₂N(R)₂, —N(R)C(O)OR,        —N(R)C(O)R, —N(R)C(S)R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂,        —N(COR)COR, —N(OR)R, —C(═NH) N(R)₂, —C(O)N(OR)R, —C(═NOR)R,        —OP(O)(OR)₂, —P(O)(R)₂, —P(O)(OR)₂, and —P(O)(H)(OR);-   each R⁶ is independently —H or —(C1-C6)alkyl;-   each R⁷ is independently —H or —(C1-C6)alkyl;-   each R⁸ is independently —(C1-C6)alkyl, —(C3-C10)-cycloalkyl,    (C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each    occurrence of R⁸ is independently substituted with 0-5 R′;-   each R¹⁰ is independently —(C3-C10)-cycloalkyl, 3- to 10-membered    heterocyclyl-, (C6-C10)-aryl, or 5- to 10-membered heteroaryl,    wherein each occurrence of R¹⁰ is independently substituted with 0-5    R′;-   each R is independently selected from:    -   H—,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl-,    -   (C3-C10)-cycloalkenyl-,    -   [(C3-C10)-cycloalkyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkyl]-O—(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-O—(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C6-C10)-aryl-O—(C1-C12)aliphatic-,    -   (C6-C10)-aryl-N(R″)—(C1-C12)aliphatic-,    -   3- to 10-membered heterocyclyl-,    -   (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-N(R″)—(C1-C12)aliphatic-,    -   5- to 10-membered heteroaryl-,    -   (5- to 10-membered heteroaryl)-(C1-C12)-aliphatic-,    -   (5- to 10-membered heteroaryl)-O—(C1-C12)-aliphatic-; and    -   (5- to 10-membered heteroaryl)-N(R″)—(C1-C12)-aliphatic-;-   wherein said heterocyclyl has 1-4 heteroatoms independently selected    from N, NH, O, S, SO, and SO₂, and said heteroaryl has 1-4    heteroatoms independently selected from N, NH, O, and S;-   wherein each occurrence of R is independently substituted with 0-5    R′;-   or when two R groups bound to the same atom, the two R groups may be    taken together with the atom to which they are bound to form a 3- to    10-membered aromatic or non-aromatic ring having 0-4 heteroatoms    independently selected from N, NH, O, S, SO, and SO₂, wherein said    ring is optionally substituted with 0-5 R′, and wherein said ring is    optionally fused to a (C6-C10)aryl, 5- to 10-membered heteroaryl,    (C3-C10)cycloalkyl, or a 3- to 10-membered heterocyclyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl,    5- to 10-membered heteroaryl-, (C6-C10)-aryl-, (5- to 10-membered    heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to    10-membered heteroaryl)-O—(C1-C6)-alkyl-, and    (C6-C10)-aryl-O—(C1-C6)-alkyl-.

Some embodiments of this application provide a compound of formula I:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   U and the two carbon atoms designated by α and β together form a 5-    or 6-membered aromatic ring having 0-2 nitrogen atoms;-   A is C, CR⁶, or N;-   B and F are each independently selected from C, CR⁶, and N, wherein    B and F cannot both be N;-   D is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   E is N, NR⁷, CR⁶ or C(R⁶)₂;-   W is N, NR⁷, CR⁶ or C(R⁶)₂;-   X is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   Y and Z are each independently selected from C, CR⁶, and N, wherein    Y and Z cannot both be N;-   V is C or CR⁶,-   or when Z is C or CR⁶, V is C, CR⁶, or N;-   wherein when the ring formed by X, Y, Z, V and W is

then R² is —OR⁸, —SR⁸, or —(CH₂)_(n)OR⁸;

-   m and n are each independently an integer selected from 0-4;-   each occurrence of the bond “    ” is either a single bond or a double bond;-   each occurrence of R¹, R², R⁴, and R⁵ are each independently    selected from: halogen, —R, —OR, —NO₂, —NCS, —CN, —CF₃, —OCF₃,    —SiR₃, —N(R)₂, —SR, —SOR, —SO₂R, —SO₂N(R)₂, —SO₃R, —(CR₂)₁₋₃R,    —(CR₂)₁₋₃—OR, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃R, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃OR,    —C(O)R, —C(O)C(O)R, —C(O)CH₂C(O)R, —C(S)R, —C(S)OR, —C(O)OR,    —C(O)C(O)OR, —C(O)C(O)N(R)₂, —OC(O)R, —C(O)N(R)₂, —OC(O)N(R)₂,    —C(S)N(R)₂, —(CR₂)₀₋₃NHC(O)R, —N(R)N(R)COR, —N(R)N(R)C(O)OR,    —N(R)N(R)CON(R)₂, —N(R)SO₂R, —N(R)SO₂N(R)₂, —N(R)C(O)OR, —N(R)C(O)R,    —N(R)C(S)R, —N(R)C(O)N(R)₂, —N(R)C (S)N(R)₂, —N(COR)COR, —N(OR)R,    —C(═NH)N(R)₂, —C(O)N(OR)R, —C(═NOR)R, —O P(O)(OR)₂, —P(O)(R)₂,    —P(O)(OR)₂, and —P(O)(H)(OR);-   R³ is absent or is selected from:    -   halogen, —R, —OR, —NO₂, —NCS, —CN, —CF₃, —OCF₃, —SiR₃, —N(R)₂,        —SR, —SOR, —SO₂R, —SO₂N(R)₂, —SO₃R, —(CR₂)₁₋₃R, —(CR₂)₁₋₃—OR,        —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃R, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃OR, —C(O)R,        —C(O)C(O)R, —C(O)CH₂C(O)R, —C(S)R, —C(S)OR, —C(O)OR,        —C(O)C(O)OR, —C(O)C(O)N(R)₂, —OC(O)R, —C(O)N(R)₂, —OC(O)N(R)₂,        —C(S)N(R)₂, —(CR₂)₀₋₃NHC(O)R, —N(R)N(R)COR, —N(R)N(R)C(O)OR,        —N(R)N(R)CON(R)₂, —N(R)SO₂R, —N(R)SO₂N(R)₂, —N(R)C(O)OR,        —N(R)C(O)R, —N(R)C(S)R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂,        —N(COR)COR, —N(OR)R, —C(═NH) N(R)₂, —C(O)N(OR)R, —C(═NOR)R,        —OP(O)(OR)₂, —P(O)(R)₂, —P(O)(OR)₂, and —P(O)(H)(OR);-   each R⁶ is independently —H or —(C1-C6)alkyl;-   each R⁷ is independently —H or —(C1-C6)alkyl;-   each R⁸ is independently —(C1-C6)alkyl, —(C3-C10)-cycloalkyl,    —(C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each    occurrence of R⁸ is independently substituted with 0-5 R′;-   each R is independently selected from:    -   H—,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl-,    -   (C3-C10)-cycloalkenyl-,    -   [(C3-C10)-cycloalkyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkyl]-O—(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-O—(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C6-C10)-aryl-O—(C1-C12)aliphatic-,    -   3- to 10-membered heterocyclyl-,    -   (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-,    -   5- to 10-membered heteroaryl-,    -   (5- to 10-membered heteroaryl)-(C1-C12)-aliphatic-, and    -   (5- to 10-membered heteroaryl)-O—(C1-C12)-aliphatic-;-   wherein said heterocyclyl has 1-4 heteroatoms independently selected    from N, NH, O, S, SO, and SO₂, and said heteroaryl has 1-4    heteroatoms independently selected from N, NH, O, and S;-   wherein each occurrence of R is independently substituted with 0-5    R′;-   or when two R groups bound to the same atom, the two R groups may be    taken together with the atom to which they are bound to form a 3- to    10-membered aromatic or non-aromatic ring having 0-4 heteroatoms    independently selected from N, NH, O, S, SO, and SO₂, wherein said    ring is optionally substituted with 0-5 R′, and wherein said ring is    optionally fused to a (C6-C10)aryl, 5- to 10-membered heteroaryl,    (C3-C10)cycloalkyl, or a 3- to 10-membered heterocyclyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl,    5- to 10-membered heteroaryl-, (C6-C10)-aryl-, (5- to 10-membered    heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to    10-membered heteroaryl)-O—(C1-C6)-alkyl-, and    (C6-C10)-aryl-O—(C1-C6)-alkyl-.

In another aspect, the present invention provides a compound of formulaII:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein m, R¹, R², R³, R⁴, R⁵ and R⁶    are as defined in formula I.

In another aspect, the present invention provides a compound of formulaIII:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein m, R¹, R², R³, R⁴, R⁵ and R⁶    are as defined in formula I.

In another aspect, the present invention provides a compound of formulaIV:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein R² is —OR⁸, —SR⁸, or    —(CH₂)_(n)OR⁸, wherein R² is independently substituted with 0-5 R′    and wherein m, n, R¹, R³, R⁴, R⁵, R⁶, and R⁸ are as defined in    formula I.

In another aspect, the present invention provides a compound of formulaIV:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein R² is —(CH₂)_(n)O(CH₂)_(n)R,    —(CH₂)_(p)R⁸ or —(CH₂)_(n)N(R″)R¹⁰, wherein R² is independently    substituted with 0-5 R′ and wherein m, n, p, R¹, R³, R⁴, R⁵, R⁶, R⁸,    R¹⁰, and R″ are as defined herein.

The present invention also provides pharmaceutical compositions thatcomprise a compound of formulae I, II, III, or IV, or a pharmaceuticallyacceptable salt, hydrate, solvate, polymorph, isomer, or combinationthereof.

In some embodiments, compounds of formula I are GABA_(A) α5 receptorpositive allosteric modulators. In some embodiments, compounds offormula II are GABA_(A) α5 receptor positive allosteric modulators. Insome embodiments, compounds of formula III are GABA_(A) α5 receptorpositive allosteric modulators. In some embodiments, compounds offormula IV are GABA_(A) α5 receptor positive allosteric modulators.Compounds of formula I, II, III or IV can be used to treat theconditions described herein, such as through activity as GABA_(A) α5receptor positive allosteric modulators.

In another aspect of the invention, there is provided a method fortreating cognitive impairment associated with a CNS disorder in asubject in need of treatment or at risk of said cognitive impairment,the method comprising the step of administering to said subject atherapeutically effective amount of a compound of the invention or apharmaceutically acceptable salt, hydrate, solvate, polymorph, isomer,or combination thereof. In some embodiments, the CNS disorder withcognitive impairment includes, without limitation, age-related cognitiveimpairment, Mild Cognitive Impairment (MCI), amnestic MCI (aMCI),Age-Associated Memory Impairment (AAMI), Age Related Cognitive Decline(ARCD), dementia, Alzheimer's Disease (AD), prodromal AD, post traumaticstress disorder (PTSD), schizophrenia, bipolar disorder, amyotrophiclateral sclerosis (ALS), cancer-therapy-related cognitive impairment,mental retardation, Parkinson's disease (PD), autism spectrum disorders,fragile X disorder, Rett syndrome, compulsive behavior, and substanceaddiction. In another aspect of the invention, there is provided amethod of preserving or improving cognitive function in a subject inneed thereof, the method comprising the step of administering to saidsubject a therapeutically effective amount of a compound of theinvention or a pharmaceutically acceptable salt, hydrate, solvate,polymorph, isomer, or combination thereof. In certain embodiments of theinvention, a compound of the invention or a pharmaceutically acceptablesalt, hydrate, solvate, polymorph, isomer, or combination thereof isadministered every 12 or 24 hours.

In some embodiments, the compounds and compositions of the presentinvention are for use as a medicament. In some embodiments, thecompounds and compositions of the present invention are for use intreating cognitive impairment associated with a CNS disorder in asubject in need of treatment or at risk of said cognitive impairment. Insome embodiments, the CNS disorder with cognitive impairment includes,without limitation, age-related cognitive impairment, Mild CognitiveImpairment (MCI), amnestic MCI (aMCI), Age-Associated Memory Impairment(AAMI), Age Related Cognitive Decline (ARCD), dementia, Alzheimer'sDisease (AD), prodromal AD, post traumatic stress disorder (PTSD),schizophrenia, bipolar disorder, amyotrophic lateral sclerosis (ALS),cancer-therapy-related cognitive impairment, mental retardation,Parkinson's disease (PD), autism spectrum disorders, fragile X disorder,Rett syndrome, compulsive behavior, and substance addiction.

In some embodiments, this application provides the use of a compound orcomposition described herein in the preparation of a medicament for thetreatment of cognitive impairment associated with a CNS disorder in asubject in need of treatment or at risk of said cognitive impairment. Insome embodiments, the CNS disorder with cognitive impairment includes,without limitation, age-related cognitive impairment, Mild CognitiveImpairment (MCI), amnestic MCI (aMCI), Age-Associated Memory Impairment(AAMI), Age Related Cognitive Decline (ARCD), dementia, Alzheimer'sDisease (AD), prodromal AD, post traumatic stress disorder (PTSD),schizophrenia, bipolar disorder, amyotrophic lateral sclerosis (ALS),cancer-therapy-related cognitive impairment, mental retardation,Parkinson's disease (PD), autism spectrum disorders, fragile X disorder,Rett syndrome, compulsive behavior, and substance addiction.

DETAILED DESCRIPTION OF THE FIGURES

FIG. 1 is a graph depicting the effects of administering methyl3,5-diphenylpyridazine-4-carboxylate on the spatial memory retention often aged-impaired (AI) rats in an eight-arm Radial Arm Maze (RAM) test.The black bars refer to rats treated with vehicle alone; open bars referto rats treated with methyl 3,5-diphenylpyridazine-4-carboxylate atdifferent doses; hatched bar refers to rats treated with the combinationof TB21007 and methyl 3,5-diphenylpyridazine-4-carboxylate.

FIG. 2 is a graph showing the effect of methyl3,5-diphenylpyridazine-4-carboxylate (administered intravenously) on thebinding of Ro154513 in the hippocampus and cerebellum. Methyl3,5-diphenylpyridazine-4-carboxylate blocked the binding of Ro154513 inthe hippocampus but did not affect binding of Ro15413 in the cerebellum.

FIG. 3 is a graph showing dose-dependent GABA_(A) α5 receptor occupancyby methyl 3,5-diphenylpyridazine-4-carboxylate administeredintravenously, with receptor occupancy determined either by the ratiobetween hippocampus (a region of high GABA_(A)α5 receptor density)exposure of RO 15-4513 and cerebellum (a region with low GABA_(A)α5receptor density) exposure of RO 15-4513, or by using the GABA_(A) α5selective compound L-655,708 (10 mg/kg, i.v.) to define full occupancy.

FIG. 4 is a graph showing exposure occupancy relationships for methyl3,5-diphenylpyridazine-4-carboxylate in hippocampus. Methyl3,5-diphenylpyridazine-4-carboxylate occupies about 32% of GABA_(A) α5receptors at exposures which are behaviorally active in aged-impairedrats.

FIG. 5 is a graph depicting the effect of ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylateon the spatial memory retention of ten aged-impaired (AI) rats in aneight-arm Radial Arm Maze (RAM) test. FIG. 5 shows the effect of ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylateon the spatial memory retention of ten aged-impaired (AI) rats in theRAM test, where the vehicle control was tested 3 times, and thedifferent doses of ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylatewere tested twice; In FIG. 5, black bars refer to rats treated withvehicle alone and open bars refer to rats treated with ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylateat different doses.

FIG. 6 is a graph showing the effect of ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylate(administered intravenously) on the binding of Ro154513 in thehippocampus and cerebellum. Ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylateblocked the binding of Ro154513 in the hippocampus but did not affectbinding of Ro15413 in the cerebellum.

FIG. 7 is a graph showing dose-dependent GABA_(A) α5 receptor occupancyby ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylateadministered intravenously, as calculated by the ratio betweenhippocampus (a region of high GABA_(A)α5 receptor density) exposure ofRO 15-4513 and cerebellum (a region with low GABA_(A)α5 receptordensity) exposure of RO 15-4513 to define full occupancy.

FIG. 8(A)-(C) are graphs showing the effect of 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one, as compared to vehicle dimethyl sulfoxide (DMSO), inaged-impaired rats using a Morris water maze behavioral task. FIG. 8(A)shows the escape latency (i.e., the average time in seconds rats took tofind the hidden platform in the water pool) during training in ratsreceived 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one and rats received vehicle DMSO; FIG. 8(B) shows the amount oftime spent in target annulus and opposite annulus by rats received 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one and rats received vehicle DMSO; FIG. 8(C) shows number ofcrossing in target annulus and opposite annulus by rats received 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one and rats received vehicle DMSO.

DETAILED DESCRIPTION OF THE INVENTION Definitions

Unless otherwise defined herein, scientific and technical terms used inthis application shall have the meanings that are commonly understood bythose of ordinary skill in the art. Generally, nomenclature used inconnection with, and techniques of, chemistry, cell and tissue culture,molecular biology, cell and cancer biology, neurobiology,neurochemistry, virology, immunology, microbiology, pharmacology,genetics and protein and nucleic acid chemistry, described herein, arethose well known and commonly used in the art.

The methods and techniques of the present invention are generallyperformed, unless otherwise indicated, according to conventional methodswell known in the art and as described in various general and morespecific references that are cited and discussed throughout thisspecification. See, e.g. “Principles of Neural Science,” McGraw-HillMedical, New York, N.Y. (2000); Motulsky, “Intuitive Biostatistics,”Oxford University Press, Inc. (1995); Lodish et al., “Molecular CellBiology, 4th ed.,” W. H. Freeman & Co., New York (2000); Griffiths etal., “Introduction to Genetic Analysis, 7th ed.,” W. H. Freeman & Co.,N.Y. (1999); and Gilbert et al., “Developmental Biology, 6th ed.,”Sinauer Associates, Inc., Sunderland, Mass. (2000).

Chemistry terms used herein are used according to conventional usage inthe art, as exemplified by “The McGraw-Hill Dictionary of ChemicalTerms,” Parker S., Ed., McGraw-Hill, San Francisco, C.A. (1985).

All of the publications, patents and published patent applicationsreferred to in this application are specifically incorporated byreference herein. In case of conflict, the present specification,including its specific definitions, will control.

Throughout this specification, the word “comprise” or variations such as“comprises” or “comprising” will be understood to imply the inclusion ofa stated integer (or components) or group of integers (or components),but not the exclusion of any other integer (or components) or group ofintegers (or components).

The singular forms “a,” “an,” and “the” include the plurals unless thecontext clearly dictates otherwise.

The term “including” is used to mean “including but not limited to”.“Including” and “including but not limited to” are used interchangeably.

The term “agent” is used herein to denote a chemical compound (such asan organic or inorganic compound (including, such as, a compound of thepresent invention), a mixture of chemical compounds), a biologicalmacromolecule (such as a nucleic acid, an antibody, including partsthereof as well as humanized, chimeric and human antibodies andmonoclonal antibodies, a protein or portion thereof, e.g., a peptide, alipid, a carbohydrate), or an extract made from biological materialssuch as bacteria, plants, fungi, or animal (particularly mammalian)cells or tissues. Agents include, for example, agents which are knownwith respect to structure, and those which are not known with respect tostructure. The α5-containing GABA_(A) receptor agonist activity of suchagents may render them suitable as “therapeutic agents” in the methodsand compositions of this invention.

A “patient,” “subject,” or “individual” are used interchangeably andrefer to either a human or a non-human animal. These terms includemammals, such as humans, primates, livestock animals (including bovine,porcine, etc.), companion animals (e.g., canine, feline, etc.) androdents (e.g., mice and rats).

“Cognitive function” or “cognitive status” refers to any higher orderintellectual brain process or brain state, respectively, involved inlearning and/or memory including, but not limited to, attention,information acquisition, information processing, working memory,short-term memory, long-term memory, anterograde memory, retrogradememory, memory retrieval, discrimination learning, decision-making,inhibitory response control, attentional set-shifting, delayedreinforcement learning, reversal learning, the temporal integration ofvoluntary behavior, expressing an interest in one's surroundings andself-care, speed of processing, reasoning and problem solving and socialcognition.

In humans, cognitive function may be measured, for example and withoutlimitation, by the clinical global impression of change scale(CIBIC-plus scale); the Mini Mental State Exam (MMSE); theNeuropsychiatric Inventory (NPI); the Clinical Dementia Rating Scale(CDR); the Cambridge Neuropsychological Test Automated Battery (CANTAB);the Sandoz Clinical Assessment-Geriatric (SCAG), the Buschke SelectiveReminding Test (Buschke and Fuld, 1974); the Verbal Paired Associatessubtest; the Logical Memory subtest; the Visual Reproduction subtest ofthe Wechsler Memory Scale-Revised (WMS-R) (Wechsler, 1997); the BentonVisual Retention Test, or the explicit 3-alternative forced choice task,or MATRICS consensus neuropsychological test battery. See Folstein etal., J Psychiatric Res 12: 189-98, (1975); Robbins et al., Dementia 5:266-81, (1994); Rey, L'examen clinique en psychologie, (1964); Kluger etal., J Geriatr Psychiatry Neurol 12:168-79, (1999); Marquis et al., 2002and Masur et al., 1994. Also see Buchanan, R. W., Keefe, R. S. E.,Umbricht, D., Green, M. F., Laughren, T., and Marder, S. R. (2011), TheFDA-NIMH-MATRICS guidelines for clinical trial design ofcognitive-enhancing drugs: what do we know 5 years later? Schizophr.Bull. 37, 1209-1217.

In animal model systems, cognitive function may be measured in variousconventional ways known in the art, including using a Morris Water Maze(MWM), Barnes circular maze, elevated radial arm maze, T maze or anyother mazes in which the animals use spatial information. Cognitivefunction can be assessed by reversal learning, extradimensional setshifting, conditional discrimination learning and assessments of rewardexpectancy. Other tests known in the art may also be used to assesscognitive function, such as novel object recognition and odorrecognition tasks.

Cognitive function may also be measured using imaging techniques such asPositron Emission Tomography (PET), functional magnetic resonanceimaging (fMRI), Single Photon Emission Computed Tomography (SPECT), orany other imaging technique that allows one to measure brain function.In animals, cognitive function may also be measured withelectrophysiological techniques.

“Promoting” cognitive function refers to affecting impaired cognitivefunction so that it more closely resembles the function of a normal,unimpaired subject. Cognitive function may be promoted to any detectabledegree, but in humans preferably is promoted sufficiently to allow animpaired subject to carry out daily activities of normal life at a levelof proficiency as close as possible to a normal, unimpaired subject oran age-matched normal, unimpaired subject.

In some cases, “promoting” cognitive function in a subject affected byage-related cognitive refers to affecting impaired cognitive function sothat it more closely resembles the function of an aged-matched normal,unimpaired subject, or the function of a young adult subject. Cognitivefunction of that subject may be promoted to any detectable degree, butin humans preferably is promoted sufficiently to allow an impairedsubject to carry out daily activities of normal life at a level ofproficiency close as possible to a normal, unimpaired subject or a youngadult subject or an age-matched normal unimpaired subject.

“Preserving” cognitive function refers to affecting normal or impairedcognitive function such that it does not decline or does not fall belowthat observed in the subject upon first presentation or diagnosis, ordelays such decline.

“Improving” cognitive function includes promoting cognitive functionand/or preserving cognitive function in a subject.

“Cognitive impairment” refers to cognitive function in subjects that isnot as robust as that expected in a normal, unimpaired subject. In somecases, cognitive function is reduced by about 5%, about 10%, about 30%,or more, compared to cognitive function expected in a normal, unimpairedsubject. In some cases, “cognitive impairment” in subjects affected byaged-related cognitive impairment refers to cognitive function insubjects that is not as robust as that expected in an aged-matchednormal, unimpaired subject, or the function of a young adult subject(i.e. subjects with mean scores for a given age in a cognitive test).

“Age-related cognitive impairment” refers to cognitive impairment inaged subjects, wherein their cognitive function is not as robust as thatexpected in an age-matched normal subject or as that expected in youngadult subjects. In some cases, cognitive function is reduced by about5%, about 10%, about 30%, or more, compared to cognitive functionexpected in an age-matched normal subject. In some cases, cognitivefunction is as expected in an age-matched normal subject, but reduced byabout 5%, about 10%, about 30%, about 50% or more, compared to cognitivefunction expected in a young adult subject. Age-related impairedcognitive function may be associated with Mild Cognitive Impairment(MCI) (including amnestic MCI and non-amnestic MCI), Age-AssociatedMemory Impairment (AAMI), and Age-related Cognitive Decline (ARCD).

“Cognitive impairment” associated with AD or related to AD or in ADrefers to cognitive function in subjects that is not as robust as thatexpected in subjects who have not been diagnosed AD using conventionalmethodologies and standards.

“Mild Cognitive Impairment” or “MCI” refers to a condition characterizedby isolated memory impairment unaccompanied other cognitiveabnormalities and relatively normal functional abilities. One set ofcriteria for a clinical characterization of MCI specifies the followingcharacteristics: (1) memory complaint (as reported by patient,informant, or physician), (2) normal activities of daily living (ADLs),(3) normal global cognitive function, (4) abnormal memory for age(defined as scoring more than 1.5 standard deviations below the mean fora given age), and (5) absence of indicators of dementia (as defined byDSM-IV guidelines). Petersen et al., Srch. Neurol. 56: 303-308 (1999);Petersen, “Mild cognitive impairment: Aging to Alzheimer's Disease.”Oxford University Press, N.Y. (2003). The cognitive deficit in subjectswith MCI may involve any cognition area or mental process includingmemory, language, association, attention, perception, problem solving,executive function and visuospatial skills. See, e.g., Winbald et al.,J. Intern. Med. 256:240-240, 2004; Meguro, Acta. Neurol. Taiwan.15:55-57, 2008; Ellison et al., CNS Spectr. 13:66-72, 2008, Petersen,Semin. Neurol. 27:22-31, 2007. MCI is further subdivided into amnesticMCI (aMCI) and non-amnestic MCI, characterized by the impairment (orlack thereof) of memory in particular. MCI is defined as aMCI if memoryis found to be impaired given the age and education level of thesubject. If, on the other hand, the memory of the subject is found to beintact for age and education, but other non-memory cognitive domains areimpaired, such as language, executive function, or visuospatial skills,MCI is defines an non-amnestic MCI. aMCI and non-amnestic MCI can bothbe further subdivided into single or multiple domain MCI. aMCI-singledomain refers to a condition where memory, but not other cognitive areasare impaired. aMCI-multiple domain refers to a condition where memoryand at least one other cognitive area are impaired. Non-amnestic MCI issingle domain or multiple domain dependent on whether nor not more thanone non-memory cognitive area is impaired. See, e.g., Peterson andNegash, CNS Spectr. 13:45-53, 2008.

Diagnosis of MCI usually entails an objective assessment of cognitiveimpairment, which can be garnered through the use of well-establishedneuropsychological tests, including the Mini Mental State Examination(MMSE), the Cambridge Neuropsychological Test Automated Battery (CANTAB)and individual tests such as Rey Auditory Verbal Learning Test (AVLT),Logical Memory Subtest of the revised Wechsler Memory Scale (WMS-R) andthe New York University (NYU) Paragraph Recall Test. See Folstein etal., J Psychiatric Res 12: 189-98 (1975); Robbins et al., Dementia 5:266-81 (1994); Kluger et al., J Geriatric Psychiatry Neurol 12:168-79(1999).

“Age-Associate Memory Impairment (AAMI)” refers to a decline in memorydue to aging. A patient may be considered to have AAMI if he or she isat least 50 years old and meets all of the following criteria: a) Thepatient has noticed a decline in memory performance, b) The patientperforms worse on a standard test of memory compared to young adults, c)All other obvious causes of memory decline, except normal aging, havebeen ruled out (in other words, the memory decline cannot be attributedto other causes such as a recent heart attack or head injury,depression, adverse reactions to medication, Alzheimer's disease, etc.).

“Age-Related Cognitive Decline (ARCD)” refers to declines in memory andcognitive abilities that are a normal consequence of aging in humans(e.g., Craik & Salthouse, 1992). This is also true in virtually allmammalian species. Age-Associated Memory Impairment refers to olderpersons with objective memory declines relative to their younger years,but cognitive functioning that is normal relative to their age peers(Crook et al., 1986). Age-Consistent Memory Decline is a less pejorativelabel which emphasizes that these are normal developmental changes(Crook, 1993; Larrabee, 1996), are not pathophysiological (Smith et al.,1991), and rarely progress to overt dementia (Youngjohn & Crook, 1993).The DSM-IV (1994) has codified the diagnostic classification of ARCD.

“Dementia” refers to a condition characterized by severe cognitivedeficit that interferes in normal activities of daily living. Subjectswith dementia also display other symptoms such as impaired judgment,changes in personality, disorientation, confusion, behavior changes,trouble speaking, and motor deficits. There are different types ofdementias, such as Alzheimer's disease (AD), vascular dementia, dementiawith Lewy bodies, and frontotemporal dementia.

Alzheimer's disease (AD) is characterized by memory deficits in itsearly phase. Later symptoms include impaired judgment, disorientation,confusion, behavior changes, trouble speaking, and motor deficits.Histologically, AD is characterized by beta-amyloid plaques and tanglesof protein tau.

Vascular dementia is caused by strokes. Symptoms overlap with those ofAD, but without the focus on memory impairment.

Dementia with Lewy bodies is characterized by abnormal deposits ofalpha-synuclein that form inside neurons in the brain. Cognitiveimpairment may be similar to AD, including impairments in memory andjudgment and behavior changes.

Frontotemporal dementia is characterized by gliosis, neuronal loss,superficial spongiform degeneration in the frontal cortex and/oranterior temporal lobes, and Picks' bodies. Symptoms include changes inpersonality and behavior, including a decline in social skills andlanguage expression/comprehension.

“Post traumatic stress disorder (PTSD)” refers to an anxiety disordercharacterized by an immediate or delayed response to a catastrophicevent, characterized by re-experiencing the trauma, psychic numbing oravoidance of stimuli associated with the trauma, and increased arousal.Re-experiencing phenomena include intrusive memories, flashbacks,nightmares, and psychological or physiological distress in response totrauma reminders. Such responses produce anxiety and can havesignificant impact, both chronic and acute, on a patient's quality oflife and physical and emotional health. PTSD is also associated withimpaired cognitive performance, and older individuals with PTSD havegreater decline in cognitive performance relative to control patients.

“Schizophrenia” refers to a chronic debilitating disorder, characterizedby a spectrum of psychopathology, including positive symptoms such asaberrant or distorted mental representations (e.g., hallucinations,delusions), negative symptoms characterized by diminution of motivationand adaptive goal-directed action (e.g., anhedonia, affectiveflattening, avolition), and cognitive impairment. While abnormalities inthe brain are proposed to underlie the full spectrum of psychopathologyin schizophrenia, currently available antipsychotics are largelyineffective in treating cognitive impairments in patients.

“Bipolar disorder” or “BP” or “manic depressive disorder” or “manicdepressive illness” refers to a chronic psychological/mood disorderwhich can be characterized by significant mood changes including periodsof depression and euphoric manic periods. BP may be diagnosed by askilled physician based on personal and medical history, interviewconsultation and physical examinations. The term “mania” or “manicperiods” or other variants refers to periods where an individualexhibits some or all of the following characteristics: racing thoughts,rapid speech, elevated levels of activity and agitation as well as aninflated sense of self-esteem, euphoria, poor judgment, insomnia,impaired concentration and aggression.

“Amyotrophic lateral sclerosis,” also known as ALS, refers to aprogressive, fatal, neurodegenerative disease characterized by adegeneration of motor neurons, the nerve cells in the central nervoussystem that control voluntary muscle movement. ALS is also characterizedby neuronal degeneration in the entorhinal cortex and hippocampus,memory deficits, and neuronal hyperexcitability in different brain areassuch as the cortex.

“Cancer-therapy-related cognitive impairment” refers to cognitiveimpairment that develops in subjects that are treated with cancertherapies such as chemotherapy and radiation. Cytotoxicity and otheradverse side-effects on the brain of cancer therapies result incognitive impairment in such functions as memory, learning andattention.

Parkinson's disease (PD) is a neurological disorder characterized by adecrease of voluntary movements. The afflicted patient has reduction ofmotor activity and slower voluntary movements compared to the normalindividual. The patient has characteristic “mask” face, a tendency tohurry while walking, bent over posture and generalized weakness of themuscles. There is a typical “lead-pipe” rigidity of passive movements.Another important feature of the disease is the tremor of theextremities occurring at rest and decreasing during movements.

“Autism,” as used herein, refers to an autism spectrum disordercharacterized by a neural development disorder leading to impairedsocial interaction and communication by restricted and repetitivebehavior. “Autism Spectrum Disorder” refers to a group of developmentaldisabilities that includes: autism; Asperger syndrome; pervasivedevelopmental disorder not otherwise specified (PDD-NOS or atypicalautism); Rett syndrome; and childhood disintegrative disorder.

Mental retardation is a generalized disorder characterized bysignificantly impaired cognitive function and deficits in adaptivebehaviors. Mental retardation is often defined as an IntelligenceQuotient (IQ) score of less than 70. Inborn causes are among manyunderlying causes for mental retardation. The dysfunction in neuronalcommunication is also considered one of the underlying causes for mentalretardation (Myrrhe van Spronsen and Casper C. Hoogenraad, Curr. Neurol.Neurosci. Rep. 2010, 10, 207-214).

In some instances, mental retardation includes, but are not limited to,Down syndrome, velocariofacial syndrome, fetal alcohol syndrome, FragileX syndrome, Klinefelter's syndrome, neurofibromatosis, congenitalhypothyroidism, Williams syndrome, phenylketonuria (PKU),Smith-Lemli-Opitz syndrome, Prader-Willi syndrome, Phelan-McDermidsyndrome, Mowat-Wilson syndrome, ciliopathy, Lowe syndrome and sideriumtype X-linked mental retardation. Down syndrome is a disorder thatincludes a combination of birth defects, including some degree of mentalretardation, characteristic facial features and, often, heart defects,increased infections, problems with vision and hearing, and other healthproblems. Fragile X syndrome is a prevalent form of inherited mentalretardation, occurring with a frequency of 1 in 4,000 males and 1 in8,000 females. The syndrome is also characterized by developmentaldelay, hyperactivity, attention deficit disorder, and autistic-likebehavior. There is no effective treatment for fragile X syndrome.

Obsessive compulsive disorder (“OCD”) is a mental condition that is mostcommonly characterized by intrusive, repetitive unwanted thoughts(obsessions) resulting in compulsive behaviors and mental acts that anindividual feels driven to perform (compulsion). Current epidemiologicaldata indicates that OCD is the fourth most common mental disorder in theUnited States. Some studies suggest the prevalence of OCD is between oneand three percent, although the prevalence of clinically recognized OCDis much lower, suggesting that many individuals with the disorder maynot be diagnosed. Patients with OCD are often diagnosed by apsychologist, psychiatrist, or psychoanalyst according to the Diagnosticand Statistical Manual of Mental Disorders, 4th edition text revision(DSM-IV-TR) (2000) diagnostic criteria that include characteristics ofobsessions and compulsions.

Substance addiction (e.g., drug addiction, alcohol addiction) is amental disorder. The addiction is not triggered instantaneously uponexposure to substance of abuse. Rather, it involves multiple, complexneural adaptations that develop with different time courses ranging fromhours to days to months (Kauer J. A. Nat. Rev. Neurosci. 2007, 8,844-858). The path to addiction generally begins with the voluntary useof one or more controlled substances, such as narcotics, barbiturates,methamphetamines, alcohol, nicotine, and any of a variety of other suchcontrolled substances. Over time, with extended use of the controlledsubstance(s), the voluntary ability to abstain from the controlledsubstance(s) is compromised due to the effects of prolonged use on brainfunction, and thus on behavior. As such, substance addiction generallyis characterized by compulsive substance craving, seeking and use thatpersist even in the face of negative consequences. The cravings mayrepresent changes in the underlying neurobiology of the patient whichlikely must be addressed in a meaningful way if recovery is to beobtained. Substance addiction is also characterized in many cases bywithdrawal symptoms, which for some substances are life threatening(e.g., alcohol, barbiturates) and in others can result in substantialmorbidity (which may include nausea, vomiting, fever, dizziness, andprofuse sweating), distress, and decreased ability to obtain recovery.For example, alcoholism, also known as alcohol dependence, is one suchsubstance addiction. Alcoholism is primarily characterized by foursymptoms, which include cravings, loss of control, physical dependenceand tolerance. These symptoms also may characterize addictions to othercontrolled substances. The craving for alcohol, as well as othercontrolled substances, often is as strong as the need for food or water.Thus, an alcoholic may continue to drink despite serious family, healthand/or legal ramifications.

“Treating” a condition or patient refers to taking steps to obtainbeneficial or desired results, including clinical results. Beneficial ordesired clinical results include, but are not limited to, preventing orslowing the progression of the disease or disorder, or alleviation,amelioration, or slowing the progression, of one or more symptoms ofcognitive impairment associated with CNS disorders, such as age-relatedcognitive impairment, Mild Cognitive Impairment (MCI), amnestic MCI(aMCI), Age-Associated Memory Impairment (AAMI), Age Related CognitiveDecline (ARCD), dementia, Alzheimer's Disease (AD), prodromal AD, posttraumatic stress disorder (PTSD), schizophrenia, bipolar disorder,amyotrophic lateral sclerosis (ALS), cancer-therapy-related cognitiveimpairment, mental retardation, Parkinson's disease (PD), autismspectrum disorders, fragile X disorder, Rett syndrome, compulsivebehavior, and substance addiction. In some embodiments, treatmentcomprises preventing or slowing the progression, of a CNS disorder (suchas one as described herein). In certain embodiments, treatment comprisesalleviation, amelioration, or slowing the progression of one or moresymptoms associated with that CNS disorder. In certain embodiments, thesymptom to be treated is cognitive impairment or cognitive deficit.Treating age-related cognitive impairment further comprises slowing theconversion of age-related cognitive impairment (including, but notlimited to MCI, ARCD and AAMI) into dementia (e.g., AD).

“Treating cognitive impairment” refers to taking steps to improvecognitive function in a subject with cognitive impairment so that thesubject's performance in one or more cognitive tests is improved to anydetectable degree, or is prevented from further decline. Preferably,that subject's cognitive function, after treatment of cognitiveimpairment, more closely resembles the function of a normal, unimpairedsubject. Treatment of cognitive impairment in humans may improvecognitive function to any detectable degree, but is preferably improvedsufficiently to allow the impaired subject to carry out daily activitiesof normal life at the same level of proficiency as a normal, unimpairedsubject. In some cases, “treating cognitive impairment” refers to takingsteps to improve cognitive function in a subject with cognitiveimpairment so that the subject's performance in one or more cognitivetests is improved to any detectable degree, or is prevented from furtherdecline. Preferably, that subject's cognitive function, after treatmentof cognitive impairment, more closely resembles the function of anormal, unimpaired subject. In some cases, “treating cognitiveimpairment” in a subject affecting by age-related cognitive impairmentrefers to takings steps to improve cognitive function in the subject sothat the subject's cognitive function, after treatment of cognitiveimpairment, more closely resembles the function of an age-matchednormal, unimpaired subject, or the function of a young adult subject.

“Administering” or “administration of” a substance, a compound or anagent to a subject can be carried out using one of a variety of methodsknown to those skilled in the art. For example, a compound or an agentcan be administered, intravenously, arterially, intradermally,intramuscularly, intraperitoneally, intravenously, subcutaneously,ocularly, sublingually, orally (by ingestion), intranasally (byinhalation), intraspinally, intracerebrally, and transdermally (byabsorption, e.g., through a skin duct). A compound or agent can alsoappropriately be introduced by rechargeable or biodegradable polymericdevices or other devices, e.g., patches and pumps, or formulations,which provide for the extended, slow, or controlled release of thecompound or agent. Administering can also be performed, for example,once, a plurality of times, and/or over one or more extended periods. Insome aspects, the administration includes both direct administration,including self-administration, and indirect administration, includingthe act of prescribing a drug. For example, as used herein, a physicianwho instructs a patient to self-administer a drug, or to have the drugadministered by another and/or who provides a patient with aprescription for a drug is administering the drug to the patient.

Appropriate methods of administering a substance, a compound or an agentto a subject will also depend, for example, on the age of the subject,whether the subject is active or inactive at the time of administering,whether the subject is cognitively impaired at the time ofadministering, the extent of the impairment, and the chemical andbiological properties of the compound or agent (e.g. solubility,digestibility, bioavailability, stability and toxicity). In someembodiments, a compound or an agent is administered orally, e.g., to asubject by ingestion, or intravenously, e.g., to a subject by injection.In some embodiments, the orally administered compound or agent is in anextended release or slow release formulation, or administered using adevice for such slow or extended release.

As used herein, a “α5-containing GABA_(A) receptor agonist,”“α5-containing GABA_(A) R agonist” or a “GABA_(A) α5 receptor agonist”and other variations as used herein refer to a compound that enhancesthe function of α5-containing GABA_(A) receptor (GABA_(A) R), i.e., acompound that increase GABA-gated Cl⁻ currents. In some embodiments,α5-containing GABA_(A) R agonist as used herein refers to a positiveallosteric modulator, which potentiates the activity of GABA.α5-containing GABA_(A) receptor agonists, suitable for use in thepresent invention, include the α5-containing GABA_(A) receptor agonistsof all formulas and specific α5-containing GABA_(A) receptor agonistsdescribed herein, and their hydrates, solvates, polymorphs, salts (e.g.,pharmaceutically acceptable salts), isomers (e.g., stereoisomers, E/Zisomers, and tautomers), and combinations thereof.

“Antipsychotic”, “antipsychotic agent”, “antipsychotic drug”, or“antipsychotic compound” refers to (1) a typical or an atypicalantipsychotic; (2) an agent that is selected from dopaminergic agents,glutamatergic agents, NMDA receptor positive allosteric modulators,glycine reuptake inhibitors, glutamate reuptake inhibitor, metabotropicglutamate receptors (mGluRs) agonists or positive allosteric modulators(PAMs) (e.g., mGluR2/3 agonists or PAMs), glutamate receptor glur5positive allosteric modulators (PAMs), M1 muscarinic acetylcholinereceptor (mAChR) positive allosteric modulators (PAMs), histamine H3receptor antagonists, AMPA/kainate receptor antagonists, ampakines(CX-516), glutathione prodrugs, noradrenergic agents, serotonin receptormodulators, cholinergic agents, cannabinoid CB1 antagonists, neurokinin3 antagonists, neurotensin agonists, MAO B inhibitors, PDE10 inhibitors,nNOS inhibits, neurosteroids, and neurotrophic factors, alpha-7 agonistsor positive allosteric modulators (PAMs) PAMs, serotonin 2C agonists;and/or (3) an agent that is useful in treating one or more signs orsymptoms of schizophrenia or bipolar disorder (in particular, mania).

“Typical antipsychotics”, as used herein, refer to conventionalantipsychotics, which produce antipsychotic effects as well as movementrelated adverse effects related to disturbances in the nigrostriataldopamine system. These extrapyramidal side effects (EPS) includeParkinsonism, akathisia, tardive dyskinesia and dystonia. SeeBaldessarini and Tarazi in Goodman & Gilman's The Pharmacological Basisof Therapeutics 10 Edition, 2001, pp. 485-520.

“Atypical antipsychotics”, as used herein, refer to antipsychotic drugsthat produce antipsychotic effects with little or no EPS and include,but are not limited to, aripiprazole, asenapine, clozapine, iloperidone,olanzapine, lurasidone, paliperidone, quetiapine, risperidone andziprasidone. “Atypical” antipsychotics differ from conventionalantipsychotics in their pharmacological profiles. While conventionalantipsychotics are characterized principally by D₂ dopamine receptorblockade, atypical antipsychotics show antagonist effects on multiplereceptors including the 5HT_(a) and 5HT_(c) serotonin receptors andvarying degrees of receptor affinities. Atypical antipsychotic drugs arecommonly referred to as serotonin/dopamine antagonists, reflecting theinfluential hypothesis that greater affinity for the 5HT₂ receptor thanfor the D₂ receptor underlies “atypical” antipsychotic drug action or“second generation” antipsychotic drugs. However, the atypicalantipsychotics often display side effects, including, but not limitedto, weight gain, diabetes (e.g., type II diabetes mellitus),hyperlipidemia, QTc interval prolongation, myocarditis, sexual sideeffects, extrapyramidal side effects and cataract. Thus, atypicalantipsychotics do not represent a homogeneous class, given theirdifferences in the context of both alleviation of clinical symptoms andtheir potential for inducing side effects such as the ones listed above.Further, the common side effects of the atypical antipsychotics asdescribed above often limit the antipsychotic doses that can be used forthese agents.

Memantine is chemically known as 3,5-dimethyladamantan-1-amine or3,5-dimethyltricyclo[3.3.1.1^(3,7)]decan-1-amine, which is anuncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist withmoderate affinity. The proprietary names for memantine include: Axura®and Akatinol® (Merz), Namenda® (Forest Laboratories), Ebixa® and Abixa®(Lundbeck), and Memox® (Unipharm). Memantine is approved for thetreatment of moderate to severe Alzheimer's disease (AD) in the UnitedStates at a dose of up to 28 mg/day. Derivatives or analogs ofmemantine, which include compounds that structurally or chemicallyresemble memantine, are also useful in the present invention. Suchderivatives or analogs of memantine include, but are not limited tothose compounds disclosed in U.S. Pat. Nos. 3,391,142; 4,122,193;4,273,774; and 5,061,703; U.S. Patent Application PublicationUS20040087658, US20050113458, US20060205822, US20090081259,US20090124659, and US20100227852; EP Patent Application PublicationEP2260839A2; EP Patent EP1682109B1; and PCT Application PublicationWO2005079779, all of which are incorporated herein by reference.Memantine, as used in the present invention, includes memantine and itsderivatives and analogs, as well as hydrates, polymorphs, prodrugs,salts, and solvates thereof. Memantine, as used herein, also includes acomposition comprising memantine or a derivative or an analog or apharmaceutically acceptable salt, hydrate, solvate, polymorph, orprodrug thereof, wherein the composition optionally further comprises atleast one additional therapeutic agent (such as a therapeutic agentuseful for treating a CNS disorder or cognitive impairments associatedthereof). In some embodiments, the memantine composition suitable foruse in the present invention comprises memantine and a secondtherapeutic agent that is donepezil (under the trade name Aricept).

“Acetylcholinesterase inhibitor” or “AChE-I” as used herein refers to anagent that inhibits the ability of the cholinesterase enzyme to breakdown the neurotransmitter acetylcholine, thereby increasing theconcentration and duration of acetylcholine, mainly in brain synapses orneuromuscular junctions. AChE-Is suitable for use in this applicationmay include, for example, the subcategories of (i) reversiblenon-competitive inhibitors or reversible competitive inhibitors, (ii)irreversible, and (iii) quasi-irreversible inhibitors.

The term “simultaneous administration,” as used herein, means that aα5-containing GABA_(A) receptor agonist (e.g., a α5-containing GABA_(A)receptor positive allosteric modulator) and a second therapeutic agent(e.g., an antipsychotic, memantine or an AChE-I), or theirpharmaceutically acceptable salts, hydrates, solvates, or polymorphs,are administered with a time separation of no more than about 15minutes, and in some embodiments no more than about 10 minutes. When thedrugs are administered simultaneously, the α5-containing GABA_(A)receptor agonist (e.g., an α5-containing GABA_(A) receptor positiveallosteric modulator) and a second therapeutic agent (e.g., anantipsychotic, memantine or an AChE-I), or their salts, hydrates,solvates, or polymorphs, may be contained in the same dosage (e.g., aunit dosage form comprising both the α5-containing GABA_(A) receptoragonist (e.g., an α5-containing GABA_(A) receptor positive allostericmodulator) and a second therapeutic agent (e.g., an antipsychotic,memantine or an AChE-I) or in discrete dosages (e.g., the α5-containingGABA_(A) receptor agonist (e.g., an α5-containing GABA_(A) receptorpositive allosteric modulator) or its salt, hydrate, solvate, orpolymorph is contained in one dosage form and a second therapeutic agent(e.g., an antipsychotic, memantine or an AChE-I), or its salt, hydrate,solvate, or polymorph is contained in another dosage form).

The term “sequential administration” as used herein means that theα5-containing GABA_(A) receptor agonist (e.g., a α5-containing GABA_(A)receptor positive allosteric modulator) and a second therapeutic agent(e.g., an antipsychotic, memantine or an AChE-I), or theirpharmaceutically acceptable salts, hydrates, solvates, polymorphs, areadministered with a time separation of more than about 15 minutes, andin some embodiments more than about one hour, or up to 12-24 hours.Either the α5-containing GABA_(A) receptor agonist (e.g., aα5-containing GABA_(A) receptor positive allosteric modulator) or asecond therapeutic agent (e.g., an antipsychotic, memantine or anAChE-I) may be administered first. The α5-containing GABA_(A) receptoragonist (e.g., a α5-containing GABA_(A) receptor positive allostericmodulator) and a second therapeutic agent (e.g., an antipsychotic,memantine or an AChE-I), or their salts, hydrates, solvents, orpolymorphs, for sequential administration may be contained in discretedosage forms, optionally contained in the same container or package.

A “therapeutically effective amount” of a drug or agent is an amount ofa drug or an agent that, when administered to a subject will have theintended therapeutic effect, e.g. improving cognitive function in asubject, e.g., a patient having cognitive impairment associated with aCNS disorder. The full therapeutic effect does not necessarily occur byadministration of one dose, and may occur only after administration of aseries of doses. Thus, a therapeutically effective amount may beadministered in one or more administrations. The precise effectiveamount needed for a subject will depend upon, for example, the subject'ssize, health and age, the nature and extent of the cognitive impairmentor other symptoms of the CNS disorder (such as age-related cognitiveimpairment, Mild Cognitive Impairment (MCI), dementia, Alzheimer'sDisease (AD), prodromal AD, post traumatic stress disorder (PTSD),schizophrenia, bipolar, ALS, cancer-therapy-related cognitiveimpairment, mental retardation, Parkinson's disease (PD), autismspectrum disorders, fragile X disorder, Rett syndrome, compulsivebehavior, and substance addiction), and the therapeutics or combinationof therapeutics selected for administration, and the mode ofadministration. The skilled worker can readily determine the effectiveamount for a given situation by routine experimentation.

The compounds of the present invention also include prodrugs, analogs orderivatives. The term “prodrug” is art-recognized and is intended toencompass compounds or agents which, under physiological conditions, areconverted into α5-containing GABA_(A) R positive allosteric modulators.A common method for making a prodrug is to select moieties which arehydrolyzed or metabolized under physiological conditions to provide thedesired compound or agent. In other embodiments, the prodrug isconverted by an enzymatic activity of the host animal to a GABA_(A) α5receptor positive allosteric modulator.

“Analog” is used herein to refer to a compound which functionallyresembles another chemical entity, but does not share the identicalchemical structure. For example, an analog is sufficiently similar to abase or parent compound such that it can substitute for the basecompound in therapeutic applications, despite minor structuraldifferences.

“Derivative” is used herein to refer to the chemical modification of acompound. Chemical modifications of a compound can include, for example,replacement of hydrogen by an alkyl, acyl, or amino group. Many othermodifications are also possible.

The term “aliphatic” as used herein refers to a straight chained orbranched alkyl, alkenyl or alkynyl. It is understood that alkenyl oralkynyl embodiments need at least two carbon atoms in the aliphaticchain. Aliphatic groups typically contains from 1 (or 2) to 12 carbons,such as from 1 (or 2) to 4 carbons.

The term “aryl” as used herein refers to a monocyclic or bicycliccarbocyclic aromatic ring system. Aryl as used herein includes a(C6-C12)-aryl-. For example, aryl as used herein can be a C6-C10monocyclic or C8-C12 bicyclic carbocyclic aromatic ring system. In someembodiments, aryl as used herein can be a (C6-C10)-aryl-. Phenyl (or Ph)is an example of a monocyclic aromatic ring system. Bicyclic aromaticring systems include systems wherein both rings are aromatic, e.g.,naphthyl, and systems wherein only one of the two rings is aromatic,e.g., tetralin.

The term “heterocyclic” as used herein refers to a monocyclic orbicyclic non-aromatic ring system having 1 to 4 heteroatom or heteroatomgroups selected from O, N, NH, S, SO, or SO₂ in a chemically stablearrangement. Heterocyclic as used herein includes a 3- to 12-memberedheterocyclyl-having 1-4 heteroatoms independently selected from O, N,NH, S, SO, or SO₂. For example, heterocyclic as used herein can be a 3-to 10-membered monocyclic or 8- to 12-membered bicyclic non-aromaticring system having 1 to 4 heteroatom or heteroatom groups selected fromO, N, NH, S, SO, or SO₂ in a chemically stable arrangement. In someembodiments, heterocyclic as used herein can be a 3- to 10-memberedheterocyclyl-having 1-4 heteroatoms independently selected from O, N,NH, S, SO, or SO₂. In a bicyclic non-aromatic ring system embodiment of“heterocyclyl,” one or both rings may contain said heteroatom orheteroatom groups. In another bicyclic “heterocyclyl” embodiment, one ofthe two rings may be aromatic. In yet another heterocyclic ring systemembodiment, a non-aromatic heterocyclic ring may optionally be fused toan aromatic carbocycle.

Examples of heterocyclic rings include 3-1H-benzimidazol-2-one,3-(1-alkyl)-benzimidazol-2-one, 2-tetrahydrofuranyl,3-tetrahydrofuranyl, 2-tetrahydrothiophenyl, 3-tetrahydrothiophenyl,2-morpholino, 3-morpholino, 4-morpholino, 2-thiomorpholino,3-thiomorpholino, 4-thiomorpholino, 1-pyrrolidinyl, 2-pyrrolidinyl,3-pyrrolidinyl, 1-tetrahydropiperazinyl, 2-tetrahydropiperazinyl,3-tetrahydropiperazinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl,1-pyrazolinyl, 3-pyrazolinyl, 4-pyrazolinyl, 5-pyrazolinyl,1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl,2-thiazolidinyl, 3-thiazolidinyl, 4-thiazolidinyl, 1-imidazolidinyl,2-imidazolidinyl, 4-imidazolidinyl, 5-imidazolidinyl, indolinyl,tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzothiolane,benzodithiane, and 1,3-dihydro-imidazol-2-one.

The term “heteroaryl” as used herein refers to a monocyclic or bicyclicaromatic ring system having 1 to 4 heteroatom or heteroatom groupsselected from O, N, NH or S in a chemically stable arrangement.Heteroaryl as used herein includes a 5- to 12-membered heteroaryl having1-4 heteroatoms independently selected from O, N, NH or S. In someembodiments, heteroaryl as used herein can be a 5- to 10-memberedheteroaryl having 1-4 heteroatoms independently selected from O, N, NHor S. For example, heteroaryl as used herein can be a 5- to 10-memberedmonocyclic or 8- to 12-membered bicyclic aromatic ring system having 1to 4 heteroatom or heteroatom groups selected from O, N, NH or S in oneor both rings in a chemically stable arrangement. In such a bicyclicaromatic ring system embodiment of “heteroaryl”:

-   -   both rings are aromatic; and    -   one or both rings may contain said heteroatom or heteroatom        groups.

Examples of heteroaryl rings include 2-furanyl, 3-furanyl, N-imidazolyl,2-imidazolyl, 4-imidazolyl, 5-imidazolyl, benzimidazolyl, 3-isoxazolyl,4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl,N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl,2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, pyridazinyl (e.g.,3-pyridazinyl), 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (e.g.,5-tetrazolyl), triazolyl (e.g., 2-triazolyl and 5-triazolyl), 2-thienyl,3-thienyl, benzofuryl, benzothiophenyl, indolyl (e.g., 2-indolyl),pyrazolyl (e.g., 2-pyrazolyl), isothiazolyl, 1,2,3-oxadiazolyl,1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl,1,2,3-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, purinyl,pyrazinyl, 1,3,5-triazinyl, quinolinyl (e.g., 2-quinolinyl,3-quinolinyl, 4-quinolinyl), and isoquinolinyl (e.g., 1-isoquinolinyl,3-isoquinolinyl, or 4-isoquinolinyl).

The term “cycloalkyl or cycloalkenyl” refers to a monocyclic or fused orbridged bicyclic carbocyclic ring system that is not aromatic. Forexample, cycloalkyl or cycloalkenyl as used herein can be a C3-C10monocyclic or fused or bridged C8-C12 bicyclic carbocyclic ring systemthat is not aromatic. Cycloalkenyl rings have one or more units ofunsaturation. Preferred cycloalkyl or cycloalkenyl groups includecyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl,cycloheptyl, cycloheptenyl, norbornyl, adamantyl and decalinyl.

As used herein, the carbon atom designations may have the indicatedinteger and any intervening integer. For example, the number of carbonatoms in a (C1-C4)-alkyl group is 1, 2, 3, or 4. It should be understoodthat these designation refer to the total number of atoms in theappropriate group. For example, in a (C3-C10)-heterocyclyl the totalnumber of carbon atoms and heteroatoms is 3 (as in aziridine), 4, 5, 6(as in morpholine), 7, 8, 9, or 10.

“Pharmaceutically acceptable salt” is used herein to refer to an agentor a compound according to the invention that is a therapeuticallyactive, non-toxic base and acid salt form of the compounds. The acidaddition salt form of a compound that occurs in its free form as a basecan be obtained by treating said free base form with an appropriate acidsuch as an inorganic acid, for example, a hydrohalic such ashydrochloric or hydrobromic, sulfuric, nitric, phosphoric and the like;or an organic acid, such as, for example, acetic, hydroxyacetic,propanoic, lactic, pyruvic, malonic, succinic, maleic, fumaric, malic,tartaric, citric, methanesulfonic, ethanesulfonic, benzenesulfonic,p-toluenesulfonic, cyclic, salicylic, p-aminosalicylic, pamoic and thelike. See, e.g., WO 01/062726.

Compounds containing acidic protons may be converted into theirtherapeutically active, non-toxic base addition salt form, e. g. metalor amine salts, by treatment with appropriate organic and inorganicbases. Appropriate base salt forms include, for example, ammonium salts,alkali and earth alkaline metal salts, e. g., lithium, sodium,potassium, magnesium, calcium salts and the like, salts with organicbases, e. g. N-methyl-D-glucamine, hydrabamine salts, and salts withamino acids such as, for example, arginine, lysine and the like.Conversely, said salt forms can be converted into the free forms bytreatment with an appropriate base or acid.

Compounds and their salts can be in the form of a solvate, which isincluded within the scope of the present invention. Such solvatesinclude for example hydrates, alcoholates and the like. See, e.g., WO01/062726.

As used herein, the term “hydrate” refers to a combination of water witha compound wherein the water retains its molecular state as water and iseither absorbed, adsorbed or contained within a crystal lattice of thesubstrate compound.

As used herein, the term “polymorph” refers to different crystallineforms of the same compound and other solid state molecular formsincluding pseudo-polymorphs, such as hydrates (e.g., bound water presentin the crystalline structure) and solvates (e.g., bound solvents otherthan water) of the same compound. Different crystalline polymorphs havedifferent crystal structures due to a different packing of the moleculesin the lattice. This results in a different crystal symmetry and/or unitcell parameters which directly influences its physical properties suchthe X-ray diffraction characteristics of crystals or powders. Adifferent polymorph, for example, will in general diffract at adifferent set of angles and will give different values for theintensities. Therefore X-ray powder diffraction can be used to identifydifferent polymorphs, or a solid form that comprises more than onepolymorph, in a reproducible and reliable way. Crystalline polymorphicforms are of interest to the pharmaceutical industry and especially tothose involved in the development of suitable dosage forms. If thepolymorphic form is not held constant during clinical or stabilitystudies, the exact dosage form used or studied may not be comparablefrom one lot to another. It is also desirable to have processes forproducing a compound with the selected polymorphic form in high puritywhen the compound is used in clinical studies or commercial productssince Impurities present may produce undesired toxicological effects.Certain polymorphic forms may exhibit enhanced thermodynamic stabilityor may be more readily manufactured in high purity in large quantities,and thus are more suitable for inclusion in pharmaceutical formulations.Certain polymorphs may display other advantageous physical propertiessuch as lack of hygroscopic tendencies, improved solubility, andenhanced rates of dissolution due to different lattice energies.

This application contemplates all the isomers of the compounds offormulae I-IV. “Isomer” as used herein includes optical isomers (such asstereoisomers, e.g., enantiomers and diastereoisomers), Z (zusammen) orE (entgegen) isomers, and tautomers. Many of the compounds useful in themethods and compositions of this invention have at least one stereogeniccenter in their structure. This stereogenic center may be present in a Ror a S configuration, said R and S notation is used in correspondencewith the rules described in Pure Appl. Chem. (1976), 45, 11-30. Theinvention also relates to all stereoisomeric forms such as enantiomericand diastereoisomeric forms of the compounds or mixtures thereof(including all possible mixtures of stereoisomers). See, e.g., WO01/062726. Furthermore, certain compounds which contain alkenyl groupsmay exist as Z (zusammen) or E (entgegen) isomers. In each instance, theinvention includes both mixture and separate individual isomers.Multiple substituents on a piperidinyl or the azepanyl ring can alsostand in either cis or trans relationship to each other with respect tothe plane of the piperidinyl or the azepanyl ring. Some of the compoundsmay also exist in tautomeric forms. Such forms, although not explicitlyindicated in the formulae described herein, are intended to be includedwithin the scope of the present invention. With respect to the methodsand compositions of the present invention, reference to a compound orcompounds is intended to encompass that compound in each of its possibleisomeric forms and mixtures thereof unless the particular isomeric formis referred to specifically. See, e.g., WO 01/062726.

The compounds of the invention enhance the function of α5-containingGABA_(A)R, i.e., they are α5-containing GABA_(A) R agonists (e.g.,α5-containing GABA_(A) receptor positive allosteric modulators) and arecapable of increasing GABA-gated Cl⁻ currents.

The invention further provides pharmaceutical compositions comprisingone or more compounds of the invention together with a pharmaceuticallyacceptable carrier or excipient. In some embodiments, the pharmaceuticalcompositions of this application may further comprise a secondtherapeutic agent, such as an antipsychotic, memantine or an AChE-I.

The invention further provides methods for treating cognitive impairmentassociated with said CNS disorders that are responsive to positiveallosteric modulators of α5-containing GABA_(A) receptor, e.g.,age-related cognitive impairment, Mild Cognitive Impairment (MCI),amnestic MCI (aMCI), Age-Associated Memory Impairment (AAMI), AgeRelated Cognitive Decline (ARCD), dementia, Alzheimer's Disease (AD),prodromal AD, post traumatic stress disorder (PTSD), schizophrenia,bipolar disorder, amyotrophic lateral sclerosis (ALS),cancer-therapy-related cognitive impairment, mental retardation,Parkinson's disease (PD), autism spectrum disorders, fragile X disorder,Rett syndrome, compulsive behavior, and substance addiction. In certainembodiments, the method is a method of treating the age-relatedcognitive impairment, Mild Cognitive Impairment (MCI), amnestic MCI(aMCI), Age-Associated Memory Impairment (AAMI), Age Related CognitiveDecline (ARCD), dementia, Alzheimer's Disease (AD), prodromal AD, posttraumatic stress disorder (PTSD), schizophrenia, bipolar disorder,amyotrophic lateral sclerosis (ALS), cancer-therapy-related cognitiveimpairment, mental retardation, Parkinson's disease (PD), autismspectrum disorders, fragile X disorder, Rett syndrome, compulsivebehavior, and substance addiction. In certain embodiments, treatmentcomprises preventing or slowing the progression of a CNS disorder asdescribed herein (such as those described herein). In certainembodiments, treatment comprises alleviation, amelioration, or slowingthe progression of one or more symptoms associated with the CNSdisorder. In certain embodiments, the symptom to be treated is cognitiveimpairment or cognitive deficit. In another aspect of the invention,there is provided a method of preserving or improving cognitive functionin a subject in need thereof, the method comprising the step ofadministering to said subject a therapeutically effective amount of acompound of the invention or a pharmaceutically acceptable salt,hydrate, solvate, polymorph, isomer, or combination thereof.

The various CNS disorders with cognitive impairment (e.g., age-relatedcognitive impairment, Mild Cognitive Impairment (MCI), amnestic MCI(aMCI), Age-Associated Memory Impairment (AAMI), Age Related CognitiveDecline (ARCD), dementia, Alzheimer's Disease (AD), prodromal AD, posttraumatic stress disorder (PTSD), schizophrenia, bipolar disorder,amyotrophic lateral sclerosis (ALS), cancer-therapy-related cognitiveimpairment, mental retardation, Parkinson's disease (PD), autismspectrum disorders, fragile X disorder, Rett syndrome, compulsivebehavior, and substance addiction) may have a variety of etiologies.However, the symptom of cognitive impairment in each of theabove-mentioned disorders may have overlapping causes. Thus, acomposition or method of treatment that treats cognitive impairment inone CNS disorder may also treat cognitive impairment in another.

Benzodiazepine Derivatives

The present invention provides a compound of formula I:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   U and the two carbon atoms designated by α and β together form a 5-    or 6-membered aromatic ring having 0-2 nitrogen atoms;-   A is C, CR⁶, or N;-   B and F are each independently selected from C, CR⁶, and N, wherein    B and F cannot both be N;-   D is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   E is N, NR⁷, CR⁶ or C(R⁶)₂;-   W is N, NR⁷, CR⁶ or C(R⁶)₂;-   X is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   Y and Z are each independently selected from C, CR⁶, and N, wherein    Y and Z cannot both be N;-   V is C or CR⁶,-   or when Z is C or CR⁶, V is C, CR⁶, or N;-   wherein when the ring formed by X, Y, Z, V and W is

then R² is —OR⁸, —SR⁸, —(CH₂)_(n)OR⁸, —(CH₂)_(n)O(CH₂)_(n)R⁸,—(CH₂)_(p)R⁸ and —(CH₂)_(n)N(R″)R¹⁰; and wherein R² is independentlysubstituted with 0-5 R′;

-   m and n are independently integers selected from 0-4;-   p is an integer selected from 2-4;-   each occurrence of the bond “    ” is either a single bond or a double bond;-   each occurrence of R¹, R², R⁴, and R⁵ are each independently    selected from:    -   halogen, —R, —OR, —NO₂, —NCS, —CN, —CF₃, —OCF₃, —SiR₃, —N(R)₂,        —SR, —SOR, —SO₂R, —SO₂N(R)₂, —SO₃R, —(CR₂)₁₋₃R, —(CR₂)₁₋₃—OR,        —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃R, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃OR, —C(O)R,        —C(O)C(O)R, —C(O)CH₂C(O)R, —C(S)R, —C(S)OR, —C(O)OR,        —C(O)C(O)OR, —C(O)C(O)N(R)₂, —OC(O)R, —C(O)N(R)₂, —OC(O)N(R)₂,        —C(S)N(R)₂, —(CR₂)₀₋₃NHC(O)R, —N(R)N(R)COR, —N(R)N(R)C(O)OR,        —N(R)N(R)CON(R)₂, —N(R)SO₂R, —N(R)SO₂N(R)₂, —N(R)C(O)OR,        —N(R)C(O)R, —N(R)C(S)R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂,        —N(COR)COR, —N(OR)R, —C(═NH) N(R)₂, —C(O)N(OR)R, —C(═NOR)R,        —OP(O)(OR)₂, —P(O)(R)₂, —P(O)(OR)₂, and —P(O)(H)(OR);-   R³ is absent or is selected from:    -   halogen, —R, —OR, —NO₂, —NCS, —CN, —CF₃, —OCF₃, —SiR₃, —N(R)₂,        —SR, —SOR, —SO₂R, —SO₂N(R)₂, —SO₃R, —(CR₂)₁₋₃R, —(CR₂)₁₋₃—OR,        —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃R, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃OR, —C(O)R,        —C(O)C(O)R, —C(O)CH₂C(O)R, —C(S)R, —C(S)OR, —C(O)OR,        —C(O)C(O)OR, —C(O)C(O)N(R)₂, —OC(O)R, —C(O)N(R)₂, —OC(O)N(R)₂,        —C(S)N(R)₂, —(CR₂)₀₋₃NHC(O)R, —N(R)N(R)COR, —N(R)N(R)C(O)OR,        —N(R)N(R)CON(R)₂, —N(R)SO₂R, —N(R)SO₂N(R)₂, —N(R)C(O)OR,        —N(R)C(O)R, —N(R)C(S)R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂,        —N(COR)COR, —N(OR)R, —C(═NH) N(R)₂, —C(O)N(OR)R, —C(═NOR)R,        —OP(O)(OR)₂, —P(O)(R)₂, —P(O)(OR)₂, and —P(O)(H)(OR);-   each R⁶ is independently —H or —(C1-C6)alkyl;-   each R⁷ is independently —H or —(C1-C6)alkyl;-   each R⁸ is independently —(C1-C6)alkyl, —(C3-C10)-cycloalkyl,    (C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each    occurrence of R⁸ is independently substituted with 0-5 R′;-   each R¹⁰ is independently —(C3-C10)-cycloalkyl, 3- to 10-membered    heterocyclyl-, (C6-C10)-aryl, or 5- to 10-membered heteroaryl,    wherein each occurrence of R¹⁰ is independently substituted with 0-5    R′;-   each R is independently selected from:    -   H—,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl-,    -   (C3-C10)-cycloalkenyl-,    -   [(C3-C10)-cycloalkyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkyl]-O—(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-O—(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C6-C10)-aryl-O—(C1-C12)aliphatic-,    -   (C6-C10)-aryl-N(R″)—(C1-C12)aliphatic-,    -   3- to 10-membered heterocyclyl-,    -   (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-N(R″)—(C1-C12)aliphatic-,    -   5- to 10-membered heteroaryl-,    -   (5- to 10-membered heteroaryl)-(C1-C12)-aliphatic-,    -   (5- to 10-membered heteroaryl)-O—(C1-C12)-aliphatic-; and    -   (5- to 10-membered heteroaryl)-N(R″)—(C1-C12)-aliphatic-;-   wherein said heterocyclyl has 1-4 heteroatoms independently selected    from N, NH, O, S, SO, and SO₂, and said heteroaryl has 1-4    heteroatoms independently selected from N, NH, O, and S;-   wherein each occurrence of R is independently substituted with 0-5    R′;-   or when two R groups bound to the same atom, the two R groups may be    taken together with the atom to which they are bound to form a 3- to    10-membered aromatic or non-aromatic ring having 0-4 heteroatoms    independently selected from N, NH, O, S, SO, and SO₂, wherein said    ring is optionally substituted with 0-5 R′, and wherein said ring is    optionally fused to a (C6-C10)aryl, 5- to 10-membered heteroaryl,    (C3-C10)cycloalkyl, or a 3- to 10-membered heterocyclyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;    wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl, 3- to    6-membered heterocyclyl, 5- to 10-membered heteroaryl-,    (C6-C10)-aryl-, (5- to 10-membered heteroaryl)-(C1-C6)-alkyl-,    (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to 10-membered    heteroaryl)-O—(C1-C6)-alkyl-, and (C6-C10)-aryl-O—(C1-C6)-alkyl-,    wherein each occurrence of R″ is independently substituted with 0-3    substituents selected from: halogen, —R^(∘), —OR^(∘), oxo,    —CH₂OR^(∘), —CH₂NR^(∘) ₂, —C(O)N(R^(∘))₂, —C(O)OR^(∘), —NO₂, —NCS,    —CN,    —CF₃, —OCF₃ and —N(R^(∘))₂, wherein each occurrence of R^(∘) is    independently selected from: —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl,    3- to 6-membered heterocyclyl, 5- to 10-membered heteroaryl-, and    (C6-C10)-aryl-.

In some embodiments, the present invention provides a compound offormula I:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   U and the two carbon atoms designated by α and β together form a 5-    or 6-membered aromatic ring having 0-2 nitrogen atoms;-   A is C, CR⁶, or N;-   B and F are each independently selected from C, CR⁶, and N, wherein    B and F cannot both be N;-   D is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   E is N, NR⁷, CR⁶ or C(R⁶)₂;-   W is N, NR⁷, CR⁶ or C(R⁶)₂;-   X is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   Y and Z are each independently selected from C, CR⁶, and N, wherein    Y and Z cannot both be N;-   V is C or CR⁶,-   or when Z is C or CR⁶, V is C, CR⁶, or N;-   wherein when the ring formed by X, Y, Z, V and W is

then R² is —OR⁸, —SR⁸, —(CH₂)_(n)OR⁸, —(CH₂)_(n)O(CH₂)_(n)R⁸,—(CH₂)_(p)R⁸ and —(CH₂)_(n)N(R″)R¹⁰; and wherein R² is independentlysubstituted with 0-5 R′;

-   m and n are independently integers selected from 0-4;-   p is an integer selected from 2-4;-   each occurrence of the bond “    ” is either a single bond or a double bond;-   each occurrence of R¹, R², R⁴, and R⁵ are each independently    selected from:    -   halogen, —R, —OR, —NO₂, —NCS, —CN, —CF₃, —OCF₃, —SiR₃, —N(R)₂,        —SR, —SOR, —SO₂R, —SO₂N(R)₂, —SO₃R, —(CR₂)₁₋₃R, —(CR₂)₁₋₃—OR,        —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃R, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃OR, —C(O)R,        —C(O)C(O)R, —C(O)CH₂C(O)R, —C(S)R, —C(S)OR, —C(O)OR,        —C(O)C(O)OR, —C(O)C(O)N(R)₂, —OC(O)R, —C(O)N(R)₂, —OC(O)N(R)₂,        —C(S)N(R)₂, —(CR₂)₀₋₃NHC(O)R, —N(R)N(R)COR, —N(R)N(R)C(O)OR,        —N(R)N(R)CON(R)₂, —N(R)SO₂R, —N(R)SO₂N(R)₂, —N(R)C(O)OR,        —N(R)C(O)R, —N(R)C(S)R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂,        —N(COR)COR, —N(OR)R, —C(═NH) N(R)₂, —C(O)N(OR)R, —C(═NOR)R,        —OP(O)(OR)₂, —P(O)(R)₂, —P(O)(OR)₂, and —P(O)(H)(OR);-   R³ is absent or is selected from:    -   halogen, —R, —OR, —NO₂, —NCS, —CN, —CF₃, —OCF₃, —SiR₃, —N(R)₂,        —SR, —SOR, —SO₂R, —SO₂N(R)₂, —SO₃R, —(CR₂)₁₋₃R, —(CR₂)₁₋₃—OR,        —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃R, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃OR, —C(O)R,        —C(O)C(O)R, —C(O)CH₂C(O)R, —C(S)R, —C(S)OR, —C(O)OR,        —C(O)C(O)OR, —C(O)C(O)N(R)₂, —OC(O)R, —C(O)N(R)₂, —OC(O)N(R)₂,        —C(S)N(R)₂, —(CR₂)₀₋₃NHC(O)R, —N(R)N(R)COR, —N(R)N(R)C(O)OR,        —N(R)N(R)CON(R)₂, —N(R)SO₂R, —N(R)SO₂N(R)₂, —N(R)C(O)OR,        —N(R)C(O)R, —N(R)C(S)R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂,        —N(COR)COR, —N(OR)R, —C(═NH) N(R)₂, —C(O)N(OR)R, —C(═NOR)R,        —OP(O)(OR)₂, —P(O)(R)₂, —P(O)(OR)₂, and —P(O)(H)(OR);-   each R⁶ is independently —H or —(C1-C6)alkyl;-   each R⁷ is independently —H or —(C1-C6)alkyl;-   each R⁸ is independently —(C1-C6)alkyl, —(C3-C10)-cycloalkyl,    (C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each    occurrence of R⁸ is independently substituted with 0-5 R′;-   each R¹⁰ is independently —(C3-C10)-cycloalkyl, 3- to 10-membered    heterocyclyl-, (C6-C10)-aryl, or 5- to 10-membered heteroaryl,    wherein each occurrence of R¹⁰ is independently substituted with 0-5    R′;-   each R is independently selected from:    -   H—,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl-,    -   (C3-C10)-cycloalkenyl-,    -   [(C3-C10)-cycloalkyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkyl]-O—(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-O—(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C6-C10)-aryl-O—(C1-C12)aliphatic-,    -   (C6-C10)-aryl-N(R″)—(C1-C12)aliphatic-,    -   3- to 10-membered heterocyclyl-,    -   (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-N(R″)—(C1-C12)aliphatic-,    -   5- to 10-membered heteroaryl-,    -   (5- to 10-membered heteroaryl)-(C1-C12)-aliphatic-,    -   (5- to 10-membered heteroaryl)-O—(C1-C12)-aliphatic-; and    -   (5- to 10-membered heteroaryl)-N(R″)—(C1-C12)-aliphatic-;-   wherein said heterocyclyl has 1-4 heteroatoms independently selected    from N, NH, O, S, SO, and SO₂, and said heteroaryl has 1-4    heteroatoms independently selected from N, NH, O, and S;-   wherein each occurrence of R is independently substituted with 0-5    R′;-   or when two R groups bound to the same atom, the two R groups may be    taken together with the atom to which they are bound to form a 3- to    10-membered aromatic or non-aromatic ring having 0-4 heteroatoms    independently selected from N, NH, O, S, SO, and SO₂, wherein said    ring is optionally substituted with 0-5 R′, and wherein said ring is    optionally fused to a (C6-C10)aryl, 5- to 10-membered heteroaryl,    (C3-C10)cycloalkyl, or a 3- to 10-membered heterocyclyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl,    5- to 10-membered heteroaryl-, (C6-C10)-aryl-, (5- to 10-membered    heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to    10-membered heteroaryl)-O—(C1-C6)-alkyl-, and    (C6-C10)-aryl-O—(C1-C6)-alkyl-.

Some embodiments provide a compound of formula I:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   U and the two carbon atoms designated by α and β together form a 5-    or 6-membered aromatic ring having 0-2 nitrogen atoms;-   A is C, CR⁶, or N;-   B and F are each independently selected from C, CR⁶, and N, wherein    B and F cannot both be N;-   D is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   E is N, NR⁷, CR⁶ or C(R⁶)₂;-   W is N, NR⁷, CR⁶ or C(R⁶)₂;-   X is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   Y and Z are each independently selected from C, CR⁶, and N, wherein    Y and Z cannot both be N;-   V is C or CR⁶,-   or when Z is C or CR⁶, V is C, CR⁶, or N;-   wherein when the ring formed by X, Y, Z, V and W is

then R² is —OR⁸, —SR⁸, or —(CH₂)_(n)OR⁸;

-   m and n are each independently an integer selected from 0-4;-   each occurrence of the bond “    ” is either a single bond or a double bond;-   each occurrence of R¹, R², R⁴, and R⁵ are each independently    selected from: halogen, —R, —OR, —NO₂, —NCS, —CN, —CF₃, —OCF₃,    —SiR₃, —N(R)₂, —SR, —SOR, —SO₂R, —SO₂N(R)₂, —SO₃R, —(CR₂)₁₋₃R,    —(CR₂)₁₋₃—OR, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃R, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃OR,    —C(O)R, —C(O)C(O)R, —C(O)CH₂C(O)R, —C(S)R, —C(S)OR, —C(O)OR,    —C(O)C(O)OR, —C(O)C(O)N(R)₂, —OC(O)R, —C(O)N(R)₂, —OC(O)N(R)₂,    —C(S)N(R)₂, —(CR₂)₀₋₃NHC(O)R, —N(R)N(R)COR, —N(R)N(R)C(O)OR,    —N(R)N(R)CON(R)₂, —N(R)SO₂R, —N(R)SO₂N(R)₂, —N(R)C(O)OR, —N(R)C(O)R,    —N(R)C(S)R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂, —N(COR)COR, —N(OR)R,    —C(═NH) N(R)₂, —C(O)N(OR)R, —C(═NOR)R, —OP(O)(OR)₂, —P(O)(R)₂,    —P(O)(OR)₂, and —P(O)(H)(OR);-   R³ is absent or is selected from:    -   halogen, —R, —OR, —NO₂, —NCS, —CN, —CF₃, —OCF₃, —SiR₃, —N(R)₂,        —SR, —SOR, —SO₂R, —SO₂N(R)₂, —SO₃R, —(CR₂)₁₋₃R, —(CR₂)₁₋₃—OR,        —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃R, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃OR, —C(O)R,        —C(O)C(O)R, —C(O)CH₂C(O)R, —C(S)R, —C(S)OR, —C(O)OR,        —C(O)C(O)OR, —C(O)C(O)N(R)₂, —OC(O)R, —C(O)N(R)₂, —OC(O)N(R)₂,        —C(S)N(R)₂, —(CR₂)₀₋₃NHC(O)R, —N(R)N(R)COR, —N(R)N(R)C(O)OR,        —N(R)N(R)CON(R)₂, —N(R)SO₂R, —N(R)SO₂N(R)₂, —N(R)C(O)OR,        —N(R)C(O)R, —N(R)C(S)R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂,        —N(COR)COR, —N(OR)R, —C(═NH) N(R)₂, —C(O)N(OR)R, —C(═NOR)R,        —OP(O)(OR)₂, —P(O)(R)₂, —P(O)(OR)₂, and —P(O)(H)(OR);-   each R⁶ is independently —H or —(C1-C6)alkyl;-   each R⁷ is independently —H or —(C1-C6)alkyl;-   each R⁸ is independently —(C1-C6)alkyl, —(C3-C10)-cycloalkyl,    (C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each    occurrence of R⁸ is independently substituted with 0-5 R′;-   each R is independently selected from:    -   H—,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl-,    -   (C3-C10)-cycloalkenyl-,    -   [(C3-C10)-cycloalkyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkyl]-O—(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-O—(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C6-C10)-aryl-O—(C1-C12)aliphatic-,    -   3- to 10-membered heterocyclyl-,    -   (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-,    -   5- to 10-membered heteroaryl-,    -   (5- to 10-membered heteroaryl)-(C1-C12)-aliphatic-, and    -   (5- to 10-membered heteroaryl)-O—(C1-C12)-aliphatic-;-   wherein said heterocyclyl has 1-4 heteroatoms independently selected    from N, NH, O, S, SO, and SO₂, and said heteroaryl has 1-4    heteroatoms independently selected from N, NH, O, and S;-   wherein each occurrence of R is independently substituted with 0-5    R′;-   or when two R groups bound to the same atom, the two R groups may be    taken together with the atom to which they are bound to form a 3- to    10-membered aromatic or non-aromatic ring having 0-4 heteroatoms    independently selected from N, NH, O, S, SO, and SO₂, wherein said    ring is optionally substituted with 0-5 R′, and wherein said ring is    optionally fused to a (C6-C10)aryl, 5- to 10-membered heteroaryl,    (C3-C10)cycloalkyl, or a 3- to 10-membered heterocyclyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;    wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl,    5- to 10-membered heteroaryl-, (C6-C10)-aryl-, (5- to 10-membered    heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to    10-membered heteroaryl)-O—(C1-C6)-alkyl-, and    (C6-C10)-aryl-O—(C1-C6)-alkyl-.

The present invention provides a compound of formula I:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   U and the two carbon atoms designated by α and β together form a 5-    or 6-membered aromatic ring having 0-2 nitrogen atoms;-   A is C, CR⁶, or N;-   B and F are each independently selected from C, CR⁶, and N, wherein    B and F cannot both be N;-   D is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   E is N, NR⁷, CR⁶ or C(R⁶)₂;-   W is N, NR⁷, CR⁶ or C(R⁶)₂;-   X is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   Y and Z are each independently selected from C, CR⁶, and N, wherein    Y and Z cannot both be N;-   V is C or CR⁶,-   or when Z is C or CR⁶, V is C, CR⁶, or N;-   wherein when the ring formed by X, Y, Z, V and W is

then R² is —(CH₂)_(n)OR⁸ or —(CH₂)_(n)O(CH₂)_(n)R⁸; and wherein R² isindependently substituted with 0-5 R′;

-   m and n are independently integers selected from 0-4;-   p is an integer selected from 2-4;-   each occurrence of the bond “    ” is either a single bond or a double bond;-   each R¹ is independently selected from: halogen, —R, and —OR;-   R² is selected from: halogen, —R and —(CR₂)₁₋₃—OR;-   R³ is selected from: —R and —CN;-   R⁴ and R⁵ are each independently —H or —(C1-C6)alkyl;-   each R⁶ is independently —H or —(C1-C6)alkyl;-   each R⁷ is independently —H or —(C1-C6)alkyl;-   each R⁸ is independently —(C1-C6)alkyl, —(C3-C10)-cycloalkyl,    (C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each    occurrence of R⁸ is independently substituted with 0-5 R′;-   each R is independently selected from:    -   H—,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl-,    -   (C3-C10)-cycloalkenyl-,    -   [(C3-C10)-cycloalkyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkyl]-O—(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-O—(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C6-C10)-aryl-O—(C1-C12)aliphatic-,    -   (C6-C10)-aryl-N(R″)—(C1-C12)aliphatic-,    -   3- to 10-membered heterocyclyl-,    -   (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-N(R″)—(C1-C12)aliphatic-,    -   5- to 10-membered heteroaryl-,    -   (5- to 10-membered heteroaryl)-(C1-C12)-aliphatic-,    -   (5- to 10-membered heteroaryl)-O—(C1-C12)-aliphatic-; and    -   (5- to 10-membered heteroaryl)-N(R″)—(C1-C12)-aliphatic-;-   wherein said heterocyclyl has 1-4 heteroatoms independently selected    from N, NH, O, S, SO, and SO₂, and said heteroaryl has 1-4    heteroatoms independently selected from N, NH, O, and S;-   wherein each occurrence of R is independently substituted with 0-5    R′;-   or when two R groups bound to the same atom, the two R groups may be    taken together with the atom to which they are bound to form a 3- to    10-membered aromatic or non-aromatic ring having 0-4 heteroatoms    independently selected from N, NH, O, S, SO, and SO₂, wherein said    ring is optionally substituted with 0-5 R′, and wherein said ring is    optionally fused to a (C6-C10)aryl, 5- to 10-membered heteroaryl,    (C3-C10)cycloalkyl, or a 3- to 10-membered heterocyclyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl, 3- to    6-membered heterocyclyl, 5- to 10-membered heteroaryl-,    (C6-C10)-aryl-, (5- to 10-membered heteroaryl)-(C1-C6)-alkyl-,    (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to 10-membered    heteroaryl)-O—(C1-C6)-alkyl-, and (C6-C10)-aryl-O—(C1-C6)-alkyl-,    wherein each occurrence of R″ is independently substituted with 0-5    substituents selected from: halogen, —R^(∘), —OR^(∘), oxo,    —CH₂OR^(∘), —CH₂N(R^(∘))₂, —C(O)N(R^(∘))₂, —C(O)OR^(∘), —NO₂, —NCS,    —CN, —CF₃, —OCF₃ and —N(R^(∘))₂, wherein each occurrence of R^(∘) is    independently selected from: —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl,    3- to 6-membered heterocyclyl, 5- to 10-membered heteroaryl-, and    (C6-C10)-aryl-.

The present invention provides a compound of formula I:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   U and the two carbon atoms designated by α and β together form a 5-    or 6-membered aromatic ring having 0-2 nitrogen atoms;-   A is C, CR⁶, or N;-   B and F are each independently selected from C, CR⁶, and N, wherein    B and F cannot both be N;-   D is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   E is N, NR⁷, CR⁶ or C(R⁶)₂;-   W is N, NR⁷, CR⁶ or C(R⁶)₂;-   X is N, NR⁷, O, CR⁶ or C(R⁶)₂;-   Y and Z are each independently selected from C, CR⁶, and N, wherein    Y and Z cannot both be N;-   V is C or CR⁶,-   or when Z is C or CR⁶, V is C, CR⁶, or N;-   wherein when the ring formed by X, Y, Z, V and W is

then R² is —(CH₂)_(n)OR⁸ or —(CH₂)_(n)O(CH₂)R⁸, wherein each occurrenceof R⁸ is independently —(C1-C6)alkyl or (C6-C10)-aryl (e.g., phenyl),and wherein R² is independently substituted with 0-5 R′;

-   m and n are independently integers selected from 0-4 (in some    embodiments, m is 1);-   p is an integer selected from 2-4;-   each occurrence of the bond “    ” is either a single bond or a double bond;-   each R¹ is independently selected from: —Cl, —F, —OMe, and —C≡CH;-   R² is halogen, —(CR₂)₁₋₃—OR, wherein each occurrence of R is    independently selected from —H, —(C1-C6)alkyl, (C6-C10)-aryl- (e.g.,    phenyl), and (C6-C10)-aryl-(C1-C12)aliphatic- (e.g.,    phenyl-(C1-C6)alkyl-), and wherein each occurrence of R is    independently substituted with 0-5 R′;-   R³ is selected from: —CN, —C≡CH, —C≡C—(C1-C6)alkyl, —C≡C-phenyl,

wherein R³ is substituted with 0-5 R′;

-   each occurrence of R⁴ and R⁵ is independently —H or —(C1-C6)alkyl;-   each R⁶ is independently —H or —(C1-C6)alkyl;-   each R⁷ is independently —H or —(C1-C6)alkyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl, 3- to    6-membered heterocyclyl, 5- to 10-membered heteroaryl-,    (C6-C10)-aryl-, (5- to 10-membered heteroaryl)-(C1-C6)-alkyl-,    (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to 10-membered    heteroaryl)-O—(C1-C6)-alkyl-, and (C6-C10)-aryl-O—(C1-C6)-alkyl-,    wherein each occurrence of R″ is independently substituted with 0-5    substituents selected from: halogen, —R^(∘), —OR^(∘), oxo,    —CH₂OR^(∘), —CH₂NR^(∘) ₂, —C(O)N(R^(∘))₂, —C(O)OR^(∘), —NO₂, —NCS,    —CN, —CF₃, —OCF₃ and —N(R^(∘))₂, wherein each occurrence of R^(∘) is    independently selected from: —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl,    3- to 6-membered heterocyclyl, 5- to 10-membered heteroaryl-, and    (C6-C10)-aryl-.

In some of the above embodiments, R³ is selected from:

wherein each occurrence of R″ is independently selected from—(C1-C6)-alkyl (e.g., linear or branched), —C≡CH, phenyl, thiophene, (5-to 10-membered heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-,wherein each R″ is independently substituted with 0-3 substituentsselected from: halogen, —R^(∘), —OR^(∘), oxo, —CH₂OR^(∘), —CH₂NR^(∘) ₂,—C(O)N(R^(∘))₂, —C(O)OR^(∘), —NO₂, —NCS, —CN, —CF₃, —OCF₃ and—N(R^(∘))₂, wherein each occurrence of R^(∘) is independently selectedfrom: —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl, 3- to 6-memberedheterocyclyl, 5- to 10-membered heteroaryl-, and (C6-C10)-aryl-.

In some embodiments of a compound of formula I, X, Y, Z, V and Wtogether form a 5-membered aromatic or non-aromatic ring having 1-4nitrogen atoms, wherein said ring is substituted with 0-3 R⁶ and 0-2 R⁷.In some embodiments, X, Y, Z, V and W together form a 5-memberedaromatic ring having 1-3 nitrogen atoms, wherein said ring issubstituted with 0-2 R⁶ and 0-1 R⁷.

In certain embodiments, X, Y, Z, V and W form a ring that is selectedfrom:

In some embodiments, X, Y, Z, V and W form a ring that is selected from:

In some embodiments of a compound of formula I, W is N. In someembodiments, W is N, and X, Y, Z, V and W form a ring that is selectedfrom:

In some embodiments, W is N, and X, Y, Z, V and W form a ring that isselected from:

In certain embodiments of a compound of formula I, the ring formed by X,Y, Z, V and W is.

In certain embodiments of a compound of formula I, the ring formed by X,Y, Z, V and W is:

In certain embodiments of a compound of formula I, the ring formed by X,Y, Z, V and W is selected from:

In certain embodiments of a compound of formula I, the ring formed by X,Y, Z, V and W is selected from:

In some embodiments, the ring formed by X, Y, Z, V and W is:

In some embodiments, the ring formed by X, Y, Z, V and W is:

In some embodiments of a compound of formula I, A, B, D, E and Ftogether form a 5-membered aromatic or non-aromatic ring having 1-4nitrogen atoms, wherein said ring is substituted with 0-3 R⁶ and 0-2 R⁷.In certain embodiments, A, B, D, E and F together form a 5-memberedaromatic ring having 1-3 nitrogen atoms, wherein said ring issubstituted with 0-2 R⁶ and 0-1 R⁷.

In some embodiments of a compound of formula I, A, B, D, E and F form aring that is selected from:

In certain embodiments of a compound of formula I, the ring formed by A,B, D, F and E is:

In some embodiments of a compound of formula I, the compound has astructure of formula II:

or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,isomer, or combination thereof, wherein m, R¹, R², R³, R⁴, R⁵ and R⁶ areas defined in formula I.

In some embodiments of a compound of formula I, the compound has astructure of formula III:

or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,isomer, or combination thereof, wherein m, R¹, R², R³, R⁴, R⁵ and R⁶ areas defined in formula I.

In some embodiments of a compound of formula I, the compound has astructure of formula IV:

or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,isomer, or combination thereof, wherein R² is —OR⁸, —SR⁸, or—(CH₂)_(n)OR⁸, wherein R² is independently substituted with 0-5 R′ andwherein m, n, R¹, R³, R⁴, R⁵, R⁶, and R⁸ are as defined in formula I. Insome embodiments, R² is —OR⁸. In some embodiments, R² is —(CH₂)_(n)OR⁸.

In some embodiments of a compound of formula I, the compound has astructure of formula IV:

or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,isomer, or combination thereof, wherein R² is —(CH₂)_(n)O(CH₂)_(n)R⁸,—(CH₂)_(p)R⁸ or —(CH₂)_(n)N(R″)R¹⁰, wherein R² is independentlysubstituted with 0-5 R′ and wherein m, n, p, R¹, R³, R⁴, R⁵, R⁶, R⁸,R¹⁰, and R″ are as defined herein. In some embodiments, R² is—(CH₂)_(n)O(CH₂)_(n)R⁸.

In some embodiments of a compound of formula I, II, III, or IV, eachoccurrence of R¹ is selected from: halogen, —R, —OR, —NO₂, —CN, —CF₃,—OCF₃, —N(R)₂, and —N(R)SO₂R, wherein each occurrence of R isindependently substituted with 0-5 R′. In some embodiments, eachoccurrence of R¹ is independently selected from: halogen, —H,—(C1-C6)alkyl, —OH, —O((C1-C6)alkyl), —NO₂, —CN, —CF₃, —OCF₃, —NH₂,—N((C1-C6)alkyl)₂, —N((C1-C6)alkyl)SO₂((C1-C6)alkyl), and—NHSO₂((C1-C6)alkyl), wherein said alkyl is independently substitutedwith 0-5 R′. In certain embodiments, each occurrence of R¹ isindependently selected from: —H, —F, —Cl, —Br, —OH, -Me, -Et, —OMe,—OEt, —NO₂, —CN, —CF₃, —OCF₃, —NH₂, —NMe₂, —NEt₂, —NHSO₂Me, and—NHSO₂Et. In certain embodiments of a compound of any one of formulaeI-IV, at least one R¹ is —OR. In some embodiments, the at least one R¹is —O((C1-C6)alkyl), such as —OMe.

In some embodiments of a compound of formula I, II or III, R² isselected from: halogen, —R, —OR, —NO₂, —(CR₂)₁₋₃R, —(CR₂)₁₋₃—OR, —CN,—CF₃, —C(O)NR₂, —C(O)OR, and —OCF₃, wherein each occurrence of R isindependently substituted with 0-5 R′. In some embodiments, R² isselected from:

—H, —(C1-C6)alkyl, —CH₂—O((C1-C6)alkyl),—(C((C1-C6)alkyl)₂)₁₋₃—O((C1-C6)alkyl), —OH, —O((C1-C6)alkyl), —NO₂,—CN, —CF₃, —OCF₃, (C3-C10)-cycloalkyl-, —C(O)N((C1-C6)alkyl)₂,—C(O)O((C1-C6)alkyl), 3- to 10-membered heterocyclyl-, (C6-C10)aryl-, 5-to 10-membered heteroaryl-, (C6-C10)aryl-(C1-C12)aliphatic-,(C6-C10)aryl-O—(C1-C12)aliphatic-,(C6-C10)aryl-N(R″)—(C1-C12)aliphatic-,(C6-C10)aryl-(C1-C12)aliphatic-O—, (5- to 10-memberedheteroaryl)-(C1-C12)-aliphatic-, (5- to 10-memberedheteroaryl)-O—(C1-C12)-aliphatic-, (5- to 10-memberedheteroaryl)-N(R″)—(C1-C12)-aliphatic-, (5- to 10-memberedheteroaryl)-(C1-C12)-aliphatic-O—, (3- to 10-memberedheterocyclyl)-(C1-C12)aliphatic-, (3- to 10-memberedheterocyclyl)-O—(C1-C12)aliphatic-, (3- to 10-memberedheterocyclyl)-N(R″)—(C1-C12)aliphatic-, and (3- to 10-memberedheterocyclyl)-(C1-C12)aliphatic-O—, wherein R² is independentlysubstituted with 0-5 R′.

In some embodiments of a compound of formula I, II or III, R² isselected from: —H, -Me, -Et, propyl, isopropyl, butyl, tert-butyl,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, —CF₃, —C(O)OMe,—C(O)OEt, —OMe, —CH₂OMe, —CH₂OEt, —CH₂OPh, —CH₂-pyrrolidine,—CH₂-morpholine, —CH₂-pyridine, and —CH₂Ph, wherein said R² issubstituted with 0-3 R′. In some embodiments of a compound of formula I,II or III, R² is -Me substituted with 0-3 R′ selected from —R″, —OR″,oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″, —NO₂, —NCS, —CN, —CF₃,—OCF₃ and —N(R″)₂, wherein R″ is independently selected from H,—(C1-C6)-alkyl, (C6-C10)-aryl-, and (C6-C10)-aryl-(C1-C6)-alkyl-. Insome embodiment, R² is -Me that is independently substituted with 0-3 R′selected from —N(Me)₂, —N(Et)₂ and —N(Me)(CH₂Ph).

In some embodiments of a compound of formula I, II or III, R² isselected from: —CH₂Ph, —CH₂CH₂Ph, -Ph, —OCH₂Ph, —CH₂OPh, —OCH₂CH₂Ph,—CH₂CH₂OPh, —CH₂-pyrrolidine, —CH₂-morpholine, —CH₂-pyridine, and —CH₂Phwherein said Ph, pyrrolidine, pyridine or morpholine is substituted with0-5 R′. In some embodiments of a compound of formula I, II or III, R² isselected from: —CH₂Ph, —CH₂CH₂Ph, -Ph, —OCH₂Ph, —CH₂OPh, —OCH₂CH₂Ph,—CH₂CH₂OPh, —CH₂-pyrrolidine, —CH₂-morpholine, —CH₂-pyridine, and—CH₂Ph, wherein said Ph, pyrrolidine, pyridine or morpholine issubstituted with 0-5 R′ independently selected from halogen,(C1-C6)-alkyl, —OH, —O((C1-C6)-alkyl), —CH₂OH, —CH₂O(C1-C6)-alkyl),—CH₂N(C1-C6)-alkyl)₂, —C(O)O(C1-C6)-alkyl), —C(O)N(C1-C6)-alkyl)₂, —NO₂,—CN, —CF₃, —OCF₃ and —N(C1-C6)-alkyl)₂. In some of the aboveembodiments, the -Ph, pyrrolidine, pyridine or morpholine of R² issubstituted with 0-5 R′ independently selected from —F, —Cl, —CN, -Me,-Et, —OMe, and —OEt. In some embodiments of a compound of formula I, IIor III, R² is —CH₂Ph, —CH₂OPh, —CH₂-pyridine, —CH₂-pyrrolidine, or—CH₂-morpholine wherein said -Ph, pyrrolidine, pyridine or morpholine issubstituted with 0-3 R′ independently selected from —F, —Cl, —CN, -Me,and —OMe.

In some embodiments of a compound of formula IV, R² is —OR⁸, —SR⁸,—(CH₂)_(n)OR⁸, —(CH₂)_(n)O(CH₂)_(n)R, —(CH₂)_(p)R⁸ or—(CH₂)_(n)N(R″)R¹⁰, wherein each R⁸ is independently —(C1-C6)alkyl,—(C3-C10)-cycloalkyl, (C6-C10)-aryl, or 5- to 10-membered heteroaryl,wherein each occurrence of R⁸ is independently substituted with 0-5 R′;n is an integer selected from 0-4; p is an integer selected from 2-4;and each R¹⁰ is independently —(C3-C10)-cycloalkyl, 3- to 10-memberedheterocyclyl-, (C6-C10)-aryl, or 5- to 10-membered heteroaryl, whereineach occurrence of R¹⁰ is independently substituted with 0-5 R′. In someembodiments, R² is OR⁸. In some embodiments, R² is OR⁸, wherein R⁸ is(C6-C10)-aryl, substituted with 0-5 R′. In some embodiments, R² is OR⁸,wherein R⁸ is (C6-C10)-aryl, substituted with 0-3 halogen (such as —F).In some embodiments, R² is —(CH₂)_(n)OR⁸ or —(CH₂)_(n)O(CH₂)_(n)R⁸. Insome embodiments, R² is —(CH₂)_(n)OR⁸ or —(CH₂)_(n)O(CH₂)R⁸, wherein R⁸is —(C1-C6)alkyl, (C6-C10)-aryl, or 5- to 10-membered heteroaryl,wherein each occurrence of R⁸ is independently substituted with 0-5 R′.

In some embodiments of a compound of formula I, II, III, or IV, R³ isselected from: halogen, —R, —CN, —CF₃, —SO₂R, —C(O)N(R)₂, —C(O)R and—C(O)OR, wherein each occurrence of R is independently substituted with0-5 R′. In some embodiments, R³ is selected from: —F, —Br, —Cl,—(C1-C6)alkyl, —CN, —C≡C, —CF₃, —SO₂((C1-C6)alkyl),—C(O)N((C1-C6)alkyl)₂, —C(O)NH₂, —C(O)((C1-C6)alkyl),—SO₂((C6-C10)-aryl), —C(O)O((C1-C6)alkyl), —(C2-C6)-alkenyl,—(C2-C6)-alkynyl, —(C6-C10)-aryl, 5- to 10-membered heteroaryl-, and 3-to 10-membered heterocyclyl-, wherein said alkyl, alkenyl, alkynyl,aryl, heteroaryl or heterocyclyl-is independently substituted with 0-5R′. In some embodiments of a compound of formula I, II, III, or IV, R³is selected from: —H, —C(O)OMe, —C(O)Et, —C(O)NMe₂, —C(O)NH₂, —C(O)OEt,—C(O)OCH₂(tert-butyl), —C(O)OCH₂CF₃, —C(O)O(isopropyl), —C(O)NEt₂,—CHF₂, —CN, —C≡C, —SO₂Me, —SO₂Et, —SO₂Ph(Me), —CF₃, —CHF₂, -Me, -Et,—Br, —Cl, —CH₂Ph,

wherein R⁹ is selected from —H, -Me, -Et, —CF₃, isopropyl, —OMe, —OEt,—O-isopropyl, —CH₂NMe₂, -tert-butyl and cyclopropyl.

In certain embodiments of a compound of formula I, II, III, or IV, R³ is—C(O)OMe or —C(O)OEt. In certain embodiments of a compound of formula I,II, III, or IV, R³

wherein R⁹ is selected from —H, -Me, -Et, —CF₃, isopropyl, —OMe, —OEt,—O-isopropyl, —CH₂NMe₂, -tert-butyl and cyclopropyl.

In some embodiments of a compound of formula I, II, III, or IV, R⁴ andR⁵ are each independently selected from —H, halogen and —R, wherein eachoccurrence of R is independently substituted with 0-5 R′, or R⁴ and R⁵may be taken together with the carbon atom to which they are bound toform a 3- to 10-membered aromatic or non-aromatic ring having 0-3additional heteroatoms independently selected from N, O, S, SO, and SO₂,wherein said ring is substituted with 0-5 R′. In some embodiments, R⁴and R⁵ are each independently selected from —H, -Me, -Et, —F, or R⁴ andR⁵ are taken together with the carbon atom to which they are bound toform a 3- to 8-membered aliphatic ring. In certain embodiments, both R⁴and R⁵ are —H.

In some embodiments, the present invention provides a compound offormula II:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0-3 (e.g., m is 1);-   each R¹ is independently selected from: —Cl, —F, —OMe, and —C≡CH;-   R² is halogen or —(CR₂)₁₋₃—OR, wherein each occurrence of R is    independently selected from —H, —(C1-C6)alkyl, (C6-C10)-aryl- (e.g.,    phenyl), and (C6-C10)-aryl-(C1-C12)aliphatic- (e.g.,    phenyl-(C1-C6)alkyl-), and wherein each occurrence of R is    independently substituted with 0-5 R′;-   R³ is selected from: —CN, —C≡CH, —C≡C—(C1-C6)alkyl, —C≡C-phenyl,

wherein R³ is substituted with 0-5 R′;each occurrence of R⁴ and R⁵ is independently —H or —(C1-C6)alkyl;each R⁶ is independently —H or —(C1-C6)alkyl;

-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl, 3- to    6-membered heterocyclyl, 5- to 10-membered heteroaryl-,    (C6-C10)-aryl-, (5- to 10-membered heteroaryl)-(C1-C6)-alkyl-,    (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to 10-membered    heteroaryl)-O—(C1-C6)-alkyl-, or (C6-C10)-aryl-O—(C1-C6)-alkyl-,    wherein each occurrence of R″ is independently substituted with 0-5    substituents selected from: halogen, —R^(∘), —OR^(∘), oxo,    —CH₂OR^(∘), —CH₂N(R^(∘))₂, —C(O)N(R^(∘))₂, —C(O)OR^(∘), —NO₂, —NCS,    —CN, —CF₃, —OCF₃ and —N(R^(∘))₂, wherein each occurrence of R^(∘) is    independently selected from: —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl,    3- to 6-membered heterocyclyl, 5- to 10-membered heteroaryl-, and    (C6-C10)-aryl-.

In some of the above embodiments, R³ is selected from:

-   wherein each occurrence of R″ is independently selected from    —(C1-C6)-alkyl (e.g., linear or branched), —C≡CH, phenyl, thiophene,    (5- to 10-membered heteroaryl)-(C1-C6)-alkyl-, and    (C6-C10)-aryl-(C1-C6)-alkyl-, wherein each R″ is independently    substituted with 0-3 substituents selected from: halogen, —R^(∘),    —OR^(∘), oxo, —CH₂OR^(∘), —CH₂N(R^(∘))₂, —C(O)N(R^(∘))₂,    —C(O)OR^(∘), —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R^(∘))₂, wherein    each occurrence of R^(∘) is independently selected from:    —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl, 3- to 6-membered    heterocyclyl, 5- to 10-membered heteroaryl-, and (C6-C10)-aryl-.

In some embodiments, the present invention provides a compound offormula II:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0-3;-   each R¹ is independently selected from: halogen (e.g., Cl, F), —H,    —(C1-C6)alkyl, —OH, —O((C1-C6)alkyl) (e.g., —OMe), —NO₂, —CN, —CF₃,    and —OCF₃, wherein R¹ is independently substituted with 0-5 R′;-   R² is selected from:    -   —H, halogen, —(C1-C6)alkyl, —OH, —O((C1-C6)alkyl),        —C(O)O((C1-C6)alkyl), —C(O)NR₂,    -   (C6-C10)-aryl- (e.g., phenyl),    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C6-C10)-aryl-O—(C1-C12)aliphatic-,    -   (C6-C10)-aryl-N(R″)—(C1-C12)aliphatic-,    -   (5- to 10-membered heteroaryl)-(C1-C12)aliphatic-,    -   (5- to 10-membered heteroaryl)-O—(C1-C12)aliphatic-,    -   (5- to 10-membered heteroaryl)-N(R″)—(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-, and    -   (3- to 10-membered heterocyclyl)-N(R″)—(C1-C12)aliphatic-,-   wherein R² is independently substituted with 0-5 R′;-   R³ is selected from:    -   (C1-C6)alkyl, —(C2-C6)alkenyl (e.g., —CH═CH₂), —C≡CH, —CN,        halogen (e.g., Br), —SO₂((C6-C10)-aryl), —SO₂((C1-C6)alkyl),        —C(O)N((C1-C6)alkyl)₂, —C(O)NH₂, —C(O)O((C1-C6)alkyl),        —C(O)((C1-C6)alkyl), —(C6-C10)aryl, 5- to 10-membered heteroaryl        (e.g., 5-membered heteroaryl such as an optionally substituted

and 5- to 10-membered heterocyclyl (e.g., 5-membered heterocyclyl suchas an optionally substituted

wherein R³ is independently substituted with 0-5 R′;

-   R⁴ and R⁵ are each independently selected from —H, halogen and    —(C1-C6)alkyl;-   R⁶ is selected from —H and —(C1-C6)alkyl;-   each R is independently selected from:    -   H—,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl-,    -   (C3-C10)-cycloalkenyl-,    -   [(C3-C10)-cycloalkyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkyl]-O—(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-O—(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C6-C10)-aryl-O—(C1-C12)aliphatic-,    -   (C6-C10)-aryl-N(R″)—(C1-C12)aliphatic-,    -   3- to 10-membered heterocyclyl-,    -   (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-N(R″)—(C1-C12)aliphatic-,    -   5- to 10-membered heteroaryl-,    -   (5- to 10-membered heteroaryl)-(C1-C12)-aliphatic-,    -   (5- to 10-membered heteroaryl)-O—(C1-C12)-aliphatic-; and    -   (5- to 10-membered heteroaryl)-N(R″)—(C1-C12)-aliphatic-;-   wherein said heterocyclyl has 1-4 heteroatoms independently selected    from N, NH, O, S, SO, and SO₂, and said heteroaryl has 1-4    heteroatoms independently selected from N, NH, O, and S;-   wherein each occurrence of R is independently substituted with 0-5    R′;-   or when two R groups bound to the same atom, the two R groups may be    taken together with the atom to which they are bound to form a 3- to    10-membered aromatic or non-aromatic ring having 0-4 heteroatoms    independently selected from N, NH, O, S, SO, and SO₂, wherein said    ring is optionally substituted with 0-5 R′, and wherein said ring is    optionally fused to a (C6-C10)aryl, 5- to 10-membered heteroaryl,    (C3-C10)cycloalkyl, or a 3- to 10-membered heterocyclyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl,    5- to 10-membered heteroaryl-, (C6-C10)-aryl-, (5- to 10-membered    heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to    10-membered heteroaryl)-O—(C1-C6)-alkyl-, and    (C6-C10)-aryl-O—(C1-C6)-alkyl-.

In some embodiments, the present invention provides a compound offormula II:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0-3;-   each R¹ is independently selected from: halogen (e.g., Cl, F), —H,    —(C1-C6)alkyl, —OH, —O((C1-C6)alkyl) (e.g., —OMe), —NO₂, —CN, —CF₃,    and —OCF₃, wherein R¹ is independently substituted with 0-5 R′;-   R² is selected from:    -   —H, —C(O)NR₂, and (C6-C10)-aryl- (e.g., phenyl);-   R³ is selected from:    -   (C1-C6)alkyl, —(C2-C6)alkenyl (e.g., —CH═CH₂), —C≡CH, —CN,        halogen (e.g., Br), —SO₂((C6-C10)-aryl), —SO₂((C1-C6)alkyl),        —C(O)N((C1-C6)alkyl)₂, —C(O)NH₂, —C(O)O((C1-C6)alkyl),        —C(O)((C1-C6)alkyl), —(C6-C10)aryl, 5- to 10-membered heteroaryl        (e.g., 5-membered heteroaryl such as an optionally substituted

and 5- to 10-membered heterocyclyl (e.g., 5-membered heterocyclyl suchas an optionally substituted

wherein R³ is independently substituted with 0-5 R′;

-   R⁴ and R⁵ are each —H, halogen and —(C1-C6)alkyl;-   R⁶ is selected from —H and —(C1-C6)alkyl;-   each R is independently selected from:    -   H—,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl-,    -   (C3-C10)-cycloalkenyl-,    -   [(C3-C10)-cycloalkyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkyl]-O—(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-O—(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C6-C10)-aryl-O—(C1-C12)aliphatic-,    -   (C6-C10)-aryl-N(R″)—(C1-C12)aliphatic-,    -   3- to 10-membered heterocyclyl-,    -   (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-N(R″)—(C1-C12)aliphatic-,    -   5- to 10-membered heteroaryl-,    -   (5- to 10-membered heteroaryl)-(C1-C12)-aliphatic-,    -   (5- to 10-membered heteroaryl)-O—(C1-C12)-aliphatic-; and    -   (5- to 10-membered heteroaryl)-N(R″)—(C1-C12)-aliphatic-;-   wherein said heterocyclyl has 1-4 heteroatoms independently selected    from N, NH, O, S, SO, and SO₂, and said heteroaryl has 1-4    heteroatoms independently selected from N, NH, O, and S;-   wherein each occurrence of R is independently substituted with 0-5    R′;-   or when two R groups bound to the same atom, the two R groups may be    taken together with the atom to which they are bound to form a 3- to    10-membered aromatic or non-aromatic ring having 0-4 heteroatoms    independently selected from N, NH, O, S, SO, and SO₂, wherein said    ring is optionally substituted with 0-5 R′, and wherein said ring is    optionally fused to a (C6-C10)aryl, 5- to 10-membered heteroaryl,    (C3-C10)cycloalkyl, or a 3- to 10-membered heterocyclyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl,    5- to 10-membered heteroaryl-, (C6-C10)-aryl-, (5- to 10-membered    heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to    10-membered heteroaryl)-O—(C1-C6)-alkyl-, and    (C6-C10)-aryl-O—(C1-C6)-alkyl-.

In some embodiments, the present invention provides a compound offormula II:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0-3;-   each R¹ is independently selected from: halogen (e.g., Cl, F) and    —O((C1-C6)alkyl) (e.g., —OMe), wherein R¹ is independently    substituted with 0-5 R′;-   R² is selected from:    -   —H, —C(O)NR₂, and (C6-C10)-aryl- (e.g., phenyl);-   R³ is selected from:    -   halogen (e.g., Br), 5- to 10-membered heteroaryl (e.g.,        5-membered heteroaryl such as an optionally substituted

and 5- to 10-membered heterocyclyl (e.g., 5-membered heterocyclyl suchas an optionally substituted

wherein R³ is independently substituted with 0-5 R′;

-   R⁴ and R⁵ are each —H;-   R⁶ is —H;-   each R is independently selected from:    -   H—,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl-,    -   (C3-C10)-cycloalkenyl-,    -   [(C3-C10)-cycloalkyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkyl]-O—(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-O—(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C6-C10)-aryl-O—(C1-C12)aliphatic-,    -   (C6-C10)-aryl-N(R″)—(C1-C12)aliphatic-,    -   3- to 10-membered heterocyclyl-,    -   (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-N(R″)—(C1-C12)aliphatic-,    -   5- to 10-membered heteroaryl-,    -   (5- to 10-membered heteroaryl)-(C1-C12)-aliphatic-,    -   (5- to 10-membered heteroaryl)-O—(C1-C12)-aliphatic-; and    -   (5- to 10-membered heteroaryl)-N(R″)—(C1-C12)-aliphatic-;-   wherein said heterocyclyl has 1-4 heteroatoms independently selected    from N, NH, O, S, SO, and SO₂, and said heteroaryl has 1-4    heteroatoms independently selected from N, NH, O, and S;-   wherein each occurrence of R is independently substituted with 0-5    R′;-   or when two R groups bound to the same atom, the two R groups may be    taken together with the atom to which they are bound to form a 3- to    10-membered aromatic or non-aromatic ring having 0-4 heteroatoms    independently selected from N, NH, O, S, SO, and SO₂, wherein said    ring is optionally substituted with 0-5 R′, and wherein said ring is    optionally fused to a (C6-C10)aryl, 5- to 10-membered heteroaryl,    (C3-C10)cycloalkyl, or a 3- to 10-membered heterocyclyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl,    5- to 10-membered heteroaryl-, (C6-C10)-aryl-, (5- to 10-membered    heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to    10-membered heteroaryl)-O—(C1-C6)-alkyl-, and    (C6-C10)-aryl-O—(C1-C6)-alkyl-.

In some embodiments, the present invention provides a compound offormula II:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0-3;-   each R¹ is independently selected from: halogen (e.g., Cl, F), —H,    —(C1-C6)alkyl, —OH, —O((C1-C6)alkyl) (e.g., —OMe), —NO₂, —CN, —CF₃,    and —OCF₃, wherein R¹ is independently substituted with 0-5 R′;-   R² is selected from:    -   —H, —(C1-C6)alkyl, —OH, —O((C1-C6)alkyl), —C(O)O((C1-C6)alkyl),        —C(O)NR₂, (C6-C10)-aryl-    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C6-C10)-aryl-O—(C1-C12)aliphatic-,    -   (C6-C10)-aryl-N(R″)—(C1-C12)aliphatic-,    -   (5- to 10-membered heteroaryl)-(C1-C12)aliphatic-,    -   (5- to 10-membered heteroaryl)-O—(C1-C12)aliphatic-,    -   (5- to 10-membered heteroaryl)-N(R″)—(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-, and    -   (3- to 10-membered heterocyclyl)-N(R″)—(C1-C12)aliphatic-,-   wherein R² is independently substituted with 0-5 R′;-   R³ is selected from:    -   (C2-C6)alkenyl (e.g., —CH═CH₂) and 5- to 10-membered        heterocyclyl (e.g., 5-membered heterocyclyl such as an        optionally substituted

wherein R³ is independently substituted with 0-5 R′;

-   R⁴ and R⁵ are each independently selected from —H, halogen and    —(C1-C6)alkyl;-   R⁶ is selected from —H and —(C1-C6)alkyl;-   each R is independently selected from:    -   H—,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl-,    -   (C3-C10)-cycloalkenyl-,    -   [(C3-C10)-cycloalkyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkyl]-O—(C1-C12)-aliphatic-,    -   [(C3-C10)-cycloalkenyl]-O—(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C6-C10)-aryl-O—(C1-C12)aliphatic-,    -   (C6-C10)-aryl-N(R″)—(C1-C12)aliphatic-,    -   3- to 10-membered heterocyclyl-,    -   (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-N(R″)—(C1-C12)aliphatic-,    -   5- to 10-membered heteroaryl-,    -   (5- to 10-membered heteroaryl)-(C1-C12)-aliphatic-,    -   (5- to 10-membered heteroaryl)-O—(C1-C12)-aliphatic-; and    -   (5- to 10-membered heteroaryl)-N(R″)—(C1-C12)-aliphatic-;-   wherein said heterocyclyl has 1-4 heteroatoms independently selected    from N, NH, O, S, SO, and SO₂, and said heteroaryl has 1-4    heteroatoms independently selected from N, NH, O, and S;-   wherein each occurrence of R is independently substituted with 0-5    R′;-   or when two R groups bound to the same atom, the two R groups may be    taken together with the atom to which they are bound to form a 3- to    10-membered aromatic or non-aromatic ring having 0-4 heteroatoms    independently selected from N, NH, O, S, SO, and SO₂, wherein said    ring is optionally substituted with 0-5 R′, and wherein said ring is    optionally fused to a (C6-C10)aryl, 5- to 10-membered heteroaryl,    (C3-C10)cycloalkyl, or a 3- to 10-membered heterocyclyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂N(R″)₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl,    5- to 10-membered heteroaryl-, (C6-C10)-aryl-, (5- to 10-membered    heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to    10-membered heteroaryl)-O—(C1-C6)-alkyl-, and    (C6-C10)-aryl-O—(C1-C6)-alkyl-.

In some embodiments, the present invention provides a compound offormula II:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0-3;-   each R¹ is independently selected from: halogen (e.g., Cl, F) and    —O((C1-C6)alkyl) (e.g., —OMe), wherein R¹ is independently    substituted with 0-5 R′;-   R² is selected from:    -   —H, —(C1-C6)alkyl,    -   (C6-C10)-aryl- (e.g., phenyl), and    -   (C6-C10)-aryl-(C1-C12)aliphatic-,-   wherein R² is independently substituted with 0-5 R′;-   R³ is selected from:    -   (C2-C6)alkenyl (e.g., —CH═CH₂) and 5- to 10-membered        heterocyclyl (e.g., 5-membered heterocyclyl such as an        optionally substituted

wherein R³ is independently substituted with 0-5 R′;

-   R⁴ and R⁵ are each —H;-   R⁶ is —H;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂N(R″)₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl,    5- to 10-membered heteroaryl-, (C6-C10)-aryl-, (5- to 10-membered    heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to    10-membered heteroaryl)-O—(C1-C6)-alkyl-, and    (C6-C10)-aryl-O—(C1-C6)-alkyl-.

In some embodiments, the present invention provides a compound offormula II:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0-3;-   each R¹ is independently selected from: halogen, —H, —(C1-C6)alkyl,    —OH, —O((C1-C6)alkyl), —NO₂, —CN, —CF₃, and —OCF₃, wherein said    alkyl is independently substituted with 0-5 R′;-   R² is selected from: —(C1-C6)alkyl, —OH, —O((C1-C6)alkyl),    —C(O)O((C1-C6)alkyl), (C6-C10)-aryl-(C1-C12)aliphatic-,    (C6-C10)-aryl-O—(C1-C12)aliphatic-,    (C6-C10)-aryl-(C1-C12)aliphatic-O—, (3- to 10-membered    heterocyclyl)-(C1-C12)aliphatic-, (5- to 10-membered    heteroaryl)-(C1-C12)-aliphatic-, (5- to 10-membered    heteroaryl)-O—(C1-C12)-aliphatic-, and (5- to 10-membered    heteroaryl)-(C1-C12)-aliphatic-O—, wherein said alkyl, aryl or    heteroaryl is independently substituted with 0-5 R′;-   R³ is selected from: —(C1-C6)alkyl, —SO₂((C1-C6)alkyl),    —C(O)N((C1-C6)alkyl)₂, and —C(O)O((C1-C6)alkyl), wherein said alkyl    is independently substituted with 0-5 R′;-   R′ is as defined herein;-   R⁴ and R⁵ are each independently selected from —H, halogen and    —(C1-C6)alkyl; and-   R⁶ is selected from —H and —(C1-C6)alkyl.

In some of the embodiments of a compound of formula II, m is 0, 1 or 2;

-   when m is 1 or 2, at least one occurrence of R¹ is halogen or    —O((C1-C6)alkyl) (such as —F and —OMe);-   R² is selected from: —(C1-C6)alkyl (e.g., -Me),    (C6-C10)-aryl-(C1-C12)aliphatic- (e.g., —CH₂Ph),    (C6-C10)-aryl-O—(C1-C12)aliphatic- (e.g., —CH₂OPh) and (3- to    10-membered heterocyclyl)-(C1-C12)aliphatic- (e.g., —CH₂-pyrrolidine    and —CH₂-morpholine), wherein said aryl (e.g., -Ph) or heterocyclyl    (e.g., pyrrolidine or morpholine) is independently substituted with    0-5 R′ independently selected from —F, -Me, and —OMe, and wherein    said alkyl (e.g., -Me) is independently substituted with 0-3 R′    selected from —N(Et)₂ and —N(Me)(CH₂Ph).    -   R³ is —C(O)O((C1-C6)alkyl) (e.g., -COOEt);-   R⁴ and R⁵ are both —H; and-   R⁶ is —H.

In some embodiments, the present invention provides a compound offormula II:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0-3;-   each R¹ is independently selected from: halogen, —H, —(C1-C6)alkyl,    —OH, —O((C1-C6)alkyl), —NO₂, —CN, —CF₃, and —OCF₃, wherein R¹ is    independently substituted with 0-5 R′;-   R² is selected from:    -   (C1-C6)alkyl, —OH, —O((C1-C6)alkyl), —C(O)O((C1-C6)alkyl),    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C6-C10)-aryl-O—(C1-C12)aliphatic-,    -   (C6-C10)-aryl-N(R″)—(C1-C12)aliphatic-,    -   (5- to 10-membered heteroaryl)-(C1-C12)aliphatic-,    -   (5- to 10-membered heteroaryl)-O—(C1-C12)aliphatic-,    -   (5- to 10-membered heteroaryl)-N(R″)—(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-,    -   (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-, and    -   (3- to 10-membered heterocyclyl)-N(R″)—(C1-C12)aliphatic-,-   wherein R² is independently substituted with 0-5 R′;-   R³ is selected from:    -   (C1-C6)alkyl, —C≡C, —CN, halogen, —SO₂((C6-C10)-aryl),        —SO₂((C1-C6)alkyl), —C(O)N((C1-C6)alkyl)₂, —C(O)NH₂,        —C(O)O((C1-C6)alkyl), —C(O)((C1-C6)alkyl), —(C6-C10)aryl, and 5-        to 10-membered heteroaryl, wherein R³ is independently        substituted with 0-5 R′;-   R⁴ and R⁵ are each independently selected from —H, halogen and    —(C1-C6)alkyl;-   R⁶ is selected from —H and —(C1-C6)alkyl; and-   R′ and R″ are as defined herein.

In some embodiments of a compound of formula II:

-   m is 0, 1 or 2;-   when m is 1 or 2, at least one occurrence of R¹ is halogen or    —O((C1-C6)alkyl);-   R² is selected from:    -   (C1-C6)alkyl, (C6-C10)-aryl-(C1-C12)aliphatic-,        (C6-C10)aryl-O—(C1-C12)aliphatic-, (5- to 10-membered        heteroaryl)-(C1-C12)aliphatic-, and (3- to 10-membered        heterocyclyl)-(C1-C12)aliphatic-, wherein R² is independently        substituted with 0-3 R¹;-   R³ is halogen, —CN, —C≡C, —C(O)NH₂, —(C1-C6)alkyl,    —C(O)((C1-C6)alkyl), —C(O)O((C1-C6)alkyl), —SO₂(Ph(Me)),

-   -   wherein R³ is independently substituted with 0-3 R′, and wherein        R⁹ is selected from —H, -Me, -Et, —CF₃, isopropyl, —OMe,        -tert-butyl, and cyclopropyl;

-   R⁴ and R⁵ are both —H;

-   R⁶ is —H; and

-   R′ is as defined herein.

In some embodiments of a compound of formula II, R³ is:

wherein R⁹ is selected from —H, -Me, -Et, —CF₃, isopropyl, —OMe, and-tert-butyl.

In some embodiments, the present invention provides a compound offormula III:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0, 1, or 2, and when m is 1 or 2, at least one occurrence of R¹    is —O((C1-C6)alkyl) (such as —OMe);-   R² is selected from: —(C1-C6)alkyl (e.g., -Me) and    (C6-C10)-aryl-(C1-C12)aliphatic- (e.g., —CH₂Ph);-   R³ is —C(O)O((C1-C6)alkyl) (e.g., -COOEt);-   R⁴ and R⁵ are both —H; and-   R⁶ is —H.

In another aspect, the present invention provides a compound of formulaIV:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0-3 (e.g., m is 1);-   each R¹ is independently selected from: —Cl, —F, —OMe, and —C≡CH;-   R² is —(CH₂)_(n)OR⁸ or —(CH₂)_(n)O(CH₂)_(n)R⁸, wherein each    occurrence of R⁸ is independently —(C1-C6)alkyl or (C6-C10)-aryl    (e.g., phenyl), and wherein R² is independently substituted with 0-5    R′;-   R³ is selected from: —CN, —C≡CH, —C≡C—(C1-C6)alkyl, —C≡C-phenyl,

wherein R³ is substituted with 0-5 R′;

-   each occurrence of R⁴ and R⁵ is independently —H or —(C1-C6)alkyl;-   each R⁶ is independently —H or —(C1-C6)alkyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl, 3- to    6-membered heterocyclyl, 5- to 10-membered heteroaryl-,    (C6-C10)-aryl-, (5- to 10-membered heteroaryl)-(C1-C6)-alkyl-,    (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to 10-membered    heteroaryl)-O—(C1-C6)-alkyl-, and (C6-C10)-aryl-O—(C1-C6)-alkyl-,    wherein each occurrence of R″ is independently substituted with 0-5    substituents selected from: halogen, —R^(∘), —OR^(∘), oxo,    —CH₂OR^(∘), —CH₂N(R^(∘))₂, —C(O)N(R^(∘))₂, —C(O)OR^(∘), —NO₂, —NCS,    —CN, —CF₃, —OCF₃ and —N(R^(∘))₂, wherein each occurrence of R^(∘) is    independently selected from: —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl,    3- to 6-membered heterocyclyl, 5- to 10-membered heteroaryl-, and    (C6-C10)-aryl-.

In some of the above embodiments, R³ is selected from:

-   wherein each occurrence of R″ is independently selected from    —(C1-C6)-alkyl (e.g., linear or branched), —C≡CH, phenyl, thiophene,    (5- to 10-membered heteroaryl)-(C1-C6)-alkyl-, and    (C6-C10)-aryl-(C1-C6)-alkyl-, wherein each R″ is independently    substituted with 0-3 substituents selected from: halogen, —R^(∘),    —OR^(∘), oxo, —CH₂OR^(∘), —CH₂N(R^(∘))₂, —C(O)N(R^(∘))₂,    —C(O)OR^(∘), —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R^(∘))₂, wherein    each occurrence of R^(∘) is independently selected from:    —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl, 3- to 6-membered    heterocyclyl, 5- to 10-membered heteroaryl-, and (C6-C10)-aryl-.

In another aspect, the present invention provides a compound of formulaIV:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0-3;-   each R¹ is independently selected from: halogen (e.g., Cl), —H,    —(C1-C6)alkyl, —C≡CH, —OH, —O((C1-C6)alkyl) (e.g., OMe), —NO₂, —CN,    —CF₃, and —OCF₃, wherein R′ is independently substituted with 0-5    R′;-   R² is selected from —OR⁸, —SR⁸, —(CH₂)_(n)OR⁸ (e.g., —CH₂OMe,    —CH₂OEt, —CH₂Oisopropyl, —CH₂Opyridyl), —(CH₂)_(n)O(CH₂)_(n)R⁸,    —(CH₂)_(p)R⁸ and —(CH₂)_(n)N(R″)R¹⁰, wherein n is an integer    selected from 0-4; p is an integer selected from 2-4; each R⁸ is    independently —(C1-C6)alkyl, —(C3-C10)-cycloalkyl, (C6-C10)-aryl, or    5- to 10-membered heteroaryl, wherein each occurrence of R⁸ is    independently substituted with 0-5 R′; each R¹⁰ is independently    —(C3-C10)-cycloalkyl, 3- to 10-membered heterocyclyl-,    (C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each    occurrence of R¹⁰ is independently substituted with 0-5 R′; and    wherein R² is independently substituted with 0-5 R′;-   R³ is selected from:    -   —H, —CN, halogen (e.g., Br), —(C1-C6)alkyl, —C≡CH,        —SO₂((C1-C6)alkyl), —C(O)N((C1-C6)alkyl)₂),        —C(O)NH((C1-C6)aliphatic)₂ (e.g., —C(O)NH((C2-C6)alkynyl)₂),        (C6-C10)-aryl-(C1-C12)aliphatic-, —C(O)((C1-C6)alkyl),        —C(O)O((C1-C6)alkyl), 5- or 6-membered heterocyclyl-(e.g.,        optionally substituted

or optionally substituted

and 5- or 6-membered heteroaryl (e.g., optionally substituted

optionally substituted,

wherein R⁹ is selected from -Me, -Et, isopropyl, —CF₃, —OMe, -OEt,—O-isopropyl, —CH₂NMe₂, and cyclopropyl; and wherein R³ is independentlysubstituted with 0-5 R′;

-   R⁴ and R⁵ are each independently selected from —H, halogen and    —(C1-C6)alkyl;-   R⁶ is selected from —H and —(C1-C6)alkyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂N(R″)₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl,    5- to 10-membered heteroaryl-, (C6-C10)-aryl-, (5- to 10-membered    heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to    10-membered heteroaryl)-O—(C1-C6)-alkyl-, and    (C6-C10)-aryl-O—(C1-C6)-alkyl-.

In another aspect, the present invention provides a compound of formulaIV:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 1;-   R¹ is —C≡CH, optionally substituted with a R′;-   R² is selected from —OR⁸, —SR⁸, —(CH₂)_(n)OR⁸ (e.g., —CH₂OMe,    —CH₂OEt, —CH₂Oisopropyl, —CH₂Opyridyl), —(CH₂)_(n)O(CH₂)_(n)R⁸,    —(CH₂)_(p)R⁸ and —(CH₂)_(n)N(R″)R¹⁰, wherein n is an integer    selected from 0-4; p is an integer selected from 2-4; each R⁸ is    independently —(C1-C6)alkyl, —(C3-C10)-cycloalkyl, (C6-C10)-aryl, or    5- to 10-membered heteroaryl, wherein each occurrence of R⁸ is    independently substituted with 0-5 R′; each R¹⁰ is independently    —(C3-C10)-cycloalkyl, 3- to 10-membered heterocyclyl-,    (C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each    occurrence of R¹⁰ is independently substituted with 0-5 R′; and    wherein R² is independently substituted with 0-5 R′;-   R³ is selected from:    -   —H, —CN, halogen (e.g., Br), —(C1-C6)alkyl, —C≡CH,        —SO₂((C1-C6)alkyl), —C(O)N((C1-C6)alkyl)₂),        —C(O)NH((C1-C6)aliphatic)₂ (e.g., —C(O)NH((C1-C6)alkynyl)₂),        (C6-C10)-aryl-(C1-C12)aliphatic-, —C(O)((C1-C6)alkyl),        —C(O)O((C1-C6)alkyl), 5- or 6-membered heterocyclyl-(e.g.,        optionally substituted

or optionally substituted

and 5- or 6-membered heteroaryl (e.g., optionally substituted

optionally substituted

wherein R⁹ is selected from -Me, -Et, isopropyl, —CF₃, —OMe, -OEt,—O-isopropyl, —CH₂NMe₂, and cyclopropyl; and wherein R³ is independentlysubstituted with 0-5 R′;

-   R⁴ and R⁵ are each independently selected from —H, halogen and    —(C1-C6)alkyl;-   R⁶ is selected from —H and —(C1-C6)alkyl;-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl,    5- to 10-membered heteroaryl-, (C6-C10)-aryl-, (5- to 10-membered    heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to    10-membered heteroaryl)-O—(C1-C6)-alkyl-, and    (C6-C10)-aryl-O—(C1-C6)-alkyl-.

In another aspect, the present invention provides a compound of formulaIV:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 1;-   each R¹ is —C≡CH, optionally substituted with a R¹;-   R² is —(CH₂)_(n)OR⁸ (e.g., —CH₂OMe, —CH₂OEt, —CH₂Oisopropyl,    —CH₂Opyridyl); and wherein R² is independently substituted with 0-5    R′;-   R³ is selected from:    -   5- or 6-membered heterocyclyl-(e.g., optionally substituted

or optionally substituted

and 5- or 6-membered heteroaryl (e.g., optionally substituted

or optionally substituted

and wherein R³ is independently substituted with 0-5 R′;

-   R⁴ and R⁵ are each —H;-   R⁶ is —H; and-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl,    5- to 10-membered heteroaryl-, (C6-C10)-aryl-, (5- to 10-membered    heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to    10-membered heteroaryl)-O—(C1-C6)-alkyl-, and    (C6-C10)-aryl-O—(C1-C6)-alkyl-.

In another aspect, the present invention provides a compound of formulaIV:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0-3;-   when m is 1 or 2, at least one occurrence of R¹ is -halogen or    —O((C1-C6)alkyl);-   each R¹ is independently selected from: halogen (e.g., Cl), —H,    —(C1-C6)alkyl, —C≡CH, —OH, —O((C1-C6)alkyl) (e.g., OMe), —NO₂, —CN,    —CF₃, and —OCF₃, wherein R¹ is independently substituted with 0-5    R′;-   R² is selected from —OR⁸, —SR⁸, —(CH₂)_(n)OR⁸ (e.g., —CH₂OMe,    —CH₂OEt, —CH₂Oisopropyl, —CH₂Opyridyl), —(CH₂)_(n)O(CH₂)_(n)R⁸,    —(CH₂)_(p)R⁸ and —(CH₂)_(n)N(R″)R¹⁰, wherein n is an integer    selected from 0-4; p is an integer selected from 2-4; each R⁸ is    independently —(C1-C6)alkyl, —(C3-C10)-cycloalkyl, (C6-C10)-aryl, or    5- to 10-membered heteroaryl, wherein each occurrence of R⁸ is    independently substituted with 0-5 R′; each R¹⁰ is independently    —(C3-C10)-cycloalkyl, 3- to 10-membered heterocyclyl-,    (C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each    occurrence of R¹⁰ is independently substituted with 0-5 R′; and    wherein R² is independently substituted with 0-5 R′;-   R³ is selected from:    -   —C≡CH, —C(O)NH((C1-C6)aliphatic)₂ (e.g.,        —C(O)NH((C1-C6)alkynyl)₂), (C6-C10)-aryl-(C1-C12)aliphatic-, 5-        or 6-membered heterocyclyl-(e.g., optionally substituted

or optionally substituted

optionally substituted

and optionally substituted

and wherein R³ is independently substituted with 0-5 R′;

-   R⁴ and R⁵ are each independently selected from —H, halogen and    —(C1-C6)alkyl;-   R⁶ is selected from —H and —(C1-C6)alkyl; and-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl,    5- to 10-membered heteroaryl-, (C6-C10)-aryl-, (5- to 10-membered    heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to    10-membered heteroaryl)-O—(C1-C6)-alkyl-, and    (C6-C10)-aryl-O—(C1-C6)-alkyl-.

In another aspect, the present invention provides a compound of formulaIV:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0-3;-   each R¹ is independently selected from: halogen (e.g., Cl), —C≡CH,    and —O((C1-C6)alkyl) (e.g., OMe), wherein R¹ is independently    substituted with 0-5 R′;-   R² is —(CH₂)_(n)OR⁸ (e.g., —CH₂OMe, —CH₂OEt, —CH₂O-isopropyl,    —CH₂O-pyridyl), wherein n is an integer selected from 0-4; R⁸ is    —(C1-C6)alkyl, —(C3-C10)-cycloalkyl, (C6-C10)-aryl, or 5- to    10-membered heteroaryl, wherein each occurrence of R⁸ is    independently substituted with 0-5 R′; and wherein R² is    independently substituted with 0-5 R′;-   R³ is selected from:    -   —C≡CH, —C(O)NH((C1-C6)aliphatic)₂ (e.g.,        —C(O)NH((C1-C6)alkynyl)₂)), (C6-C10)-aryl-(C1-C12)aliphatic-, 5-        or 6-membered heterocyclyl-(e.g., optionally substituted

or optionally substituted

optionally substituted

and optionally substituted

and wherein R³ is independently substituted with 0-5 R¹;

-   R⁴ and R⁵ are each —H;-   R⁶ is —H or —(C1-C6)alkyl; and-   wherein each occurrence of R′ is independently selected from    halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″,    —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂;-   wherein each occurrence of R″ is independently selected from H,    —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl,    5- to 10-membered heteroaryl-, (C6-C10)-aryl-, (5- to 10-membered    heteroaryl)-(C1-C6)-alkyl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to    10-membered heteroaryl)-O—(C1-C6)-alkyl-, and    (C6-C10)-aryl-O—(C1-C6)-alkyl-.

In another aspect, the present invention provides a compound of formulaIV:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0, 1, or 2, and when m is 1 or 2, at least one occurrence of R¹    is —O((C1-C6)alkyl) (such as —OMe);-   R² is OR⁸, wherein R⁸ is (C6-C10)-aryl (such as phenyl), substituted    with 0-3 halogen (such as —F);-   R³ is —C(O)O((C1-C6)alkyl) (e.g., -COOEt);-   R⁴ and R⁵ are both —H; and-   R⁶ is —H.

In another aspect, the present invention provides a compound of formulaIV:

-   or a pharmaceutically acceptable salt, hydrate, solvate, polymorph,    isomer, or combination thereof, wherein:-   m is 0-3;-   when m is 1 or 2, at least one occurrence of R¹ is -halogen or    —O((C1-C6)alkyl);-   each R¹ is independently selected from: halogen, —H, —(C1-C6)alkyl,    —OH, —O((C1-C6)alkyl), —NO₂, —CN, —CF₃, and —OCF₃, wherein R¹ is    independently substituted with 0-5 R′;-   R² is selected from —OR⁸, —SR⁸, —(CH₂)_(n)OR⁸,    —(CH₂)_(n)O(CH₂)_(n)R⁸, —(CH₂)_(p)R⁸ and —(CH₂)_(n)N(R″)R¹⁰, wherein    n is an integer selected from 0-4; p is an integer selected from    2-4; each R⁸ is independently —(C1-C6)alkyl, —(C3-C10)-cycloalkyl,    (C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each    occurrence of R⁸ is independently substituted with 0-5 R′; each R¹⁰    is independently —(C3-C10)-cycloalkyl, 3- to 10-membered    heterocyclyl-, (C6-C10)-aryl, or 5- to 10-membered heteroaryl,    wherein each occurrence of R¹⁰ is independently substituted with 0-5    R′; and wherein R² is independently substituted with 0-5 R′;-   R³ is selected from:    -   —H, —CN, halogen, —(C1-C6)alkyl, —SO₂((C1-C6)alkyl),        —C(O)N((C1-C6)alkyl)₂, —C(O)((C1-C6)alkyl),        —C(O)O((C1-C6)alkyl),

wherein R⁹ is selected from -Me, -Et, isopropyl, —CF₃, —OMe, -OEt,—O-isopropyl, —CH₂NMe₂, and cyclopropyl; and wherein R³ is independentlysubstituted with 0-5 R′;

-   R⁴ and R⁵ are each independently selected from —H, halogen and    —(C1-C6)alkyl;-   R⁶ is selected from —H and —(C1-C6)alkyl; and-   R′ and R″ are as defined herein.

In some embodiments of a compound of formula IV:

-   m is 0, 1, or 2;-   R² is —OR, —(CH₂)_(n)OR⁸, —(CH₂)_(n)O(CH₂)_(n)R⁸, wherein n is 1,    and wherein R⁸ is —(C1-C6)alkyl, (C6-C10)-aryl or 5- to 10-membered    heteroaryl, wherein R⁸ is independently substituted with 0-3 R¹;-   R³ is halogen, —H, —CN, —(C1-C6)alkyl, —C(O)((C1-C6)alkyl),    —C(O)O((C1-C6)alkyl),

wherein said alkyl is independently substituted with 0-3 R¹; R⁹ isselected from -Me, -Et, isopropyl, and —CF₃;

-   R⁴ and R⁵ are both —H;-   R⁶ is —H; and-   R′ is as defined herein.

Examples of particular compounds of the present application include:

Com- pound Structure 1

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and their pharmaceutically suitable salt, hydrate, solvate, polymorph,isomer or combination thereof.

The invention also includes various combinations of R¹, R² and R³ asdescribed above. These combinations can in turn be combined with any orall of the values of the other variables described herein. For example,R¹ can be —OR or halogen; R² can be (C1-C4)-alkyl-, —OR⁸, —(CH₂)_(n)OR⁸,or —(CH₂)_(n)O(CH₂)_(n)R⁸; and optionally R³ is —C(O)OR, or —C(O)N(R)₂.In another example, R¹ is —OR or halogen; R² is (C1-C4)-alkyl-, —OR⁸,—(CH₂)_(n)OR⁸, or —(CH₂)_(n)O(CH₂)_(n)R⁸; and R³ is a 5- or 6-memberedheteroaryl, such as

For each of above examples, compounds can have the specific values ofthe groups described herein.

Any embodiment described herein is also intended to represent unlabeledforms as well as isotopically labeled forms of the compounds, unlessotherwise indicated. Isotopically labeled compounds have structuresdepicted by the formulas given herein except that one or more atoms arereplaced by an atom having a selected atomic mass or mass number.Examples of isotopes that can be incorporated into compounds of theinvention include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorous, fluorine, and chlorine, such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N,¹⁸F, ³¹P, ³²P, ³⁵S, ³⁶Cl, ¹²⁵I respectively. The invention includesvarious isotopically labeled compounds as defined herein, for examplethose into which radioactive isotopes, such as ³H, ¹³C, and ¹⁴C, arepresent. Such isotopically labeled compounds are useful in metabolicstudies (preferably with ¹⁴C), reaction kinetic studies (with, forexample ²H or ³H), detection or imaging techniques, such as positronemission tomography (PET) or single-photon emission computed tomography(SPECT) including drug or substrate tissue distribution assays, or inradioactive treatment of patients. In particular, an ¹⁸F or labeledcompound may be particularly preferred for PET or SPECT studies.Isotopically labeled compounds of this invention and prodrugs thereofcan generally be prepared by carrying out the procedures disclosed inthe schemes or in the examples and preparations described below bysubstituting a readily available isotopically labeled reagent for anon-isotopically labeled reagent.

Any of the individual embodiments recited herein may define formula I,II, III, or IV individually or be combined to produce a preferredembodiment of this invention.

General Synthetic Methodology

The compounds of this invention may be prepared in general by methodsknown to those skilled in the art. Schemes 1-10 below provide generalsynthetic routes for the preparation of compounds of formulae I-IV.Other equivalent schemes, which will be readily apparent to the ordinaryskilled organic chemist, may alternatively be used to synthesize variousportions of the molecules as illustrated by the general schemes below.

As would be recognized by skilled practitioners, compounds of formulaeI-IV with variables other than those depicted above may be prepared byvarying chemical reagents or the synthetic routes.

Pharmaceutical Compositions and Modes of Administration

The present invention provides a pharmaceutical composition comprising apharmaceutically acceptable carrier and a compound of formulae I-IV, orpharmaceutically acceptable salts, hydrates, solvates, polymorphs,isomers, or combinations thereof.

The basic nitrogen-containing groups present in the compounds of theinvention may be quaternized with such agents as lower alkyl halides,such as methyl, ethyl, propyl, and butyl chloride, bromides and iodides;dialkyl sulfates, such as dimethyl, diethyl, dibutyl and diamylsulfates, long chain halides such as decyl, lauryl, myristyl and stearylchlorides, bromides and iodides, aralkyl halides, such as benzyl andphenethyl bromides and others. Water or oil-soluble or dispersibleproducts are thereby obtained.

It will be appreciated that compounds and agents used in thecompositions of this invention preferably should readily penetrate theblood-brain barrier when peripherally administered. Compounds whichcannot penetrate the blood-brain barrier, however, can still beeffectively administered directly into the central nervous system, e.g.,by an intraventricular or other neuro-compatible route.

In some embodiments of this invention, the α5-containing GABA_(A) Rpositive allosteric modulator is formulated with a pharmaceuticallyacceptable carrier. Pharmaceutically acceptable carriers that may beused in these compositions include, but are not limited to, ionexchangers, alumina, aluminum stearate, lecithin, serum proteins, suchas human serum albumin, buffer substances such as phosphates, glycine,sorbic acid, potassium sorbate, partial glyceride mixtures of saturatedvegetable fatty acids, water, salts or electrolytes, such as protaminesulfate, disodium hydrogen phosphate, potassium hydrogen phosphate,sodium chloride, zinc salts, colloidal silica, magnesium trisilicate,polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol,sodium carboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat. In other embodiments, no carrier is used. For example, theα5-containing GABA_(A) R agonist (e.g., a α5-containing GABA_(A)receptor positive allosteric modulator) can be administered alone or asa component of a pharmaceutical formulation (therapeutic composition).The α5-containing GABA_(A) R agonist (e.g., a α5-containing GABA_(A)receptor positive allosteric modulator) may be formulated foradministration in any convenient way for use in human medicine.

In some embodiments, the therapeutic methods of the invention includeadministering the composition of a compound or agent topically,systemically, or locally. For example, therapeutic compositions ofcompounds or agents of the invention may be formulated foradministration by, for example, injection (e.g., intravenously,subcutaneously, or intramuscularly), inhalation or insufflation (eitherthrough the mouth or the nose) or oral, buccal, sublingual, transdermal,nasal, or parenteral administration. The compositions of compounds oragents described herein may be formulated as part of an implant ordevice, or formulated for slow or extended release. When administeredparenterally, the therapeutic composition of compounds or agents for usein this invention is preferably in a pyrogen-free, physiologicallyacceptable form. Techniques and formulations generally may be found inRemington's Pharmaceutical Sciences, Meade Publishing Co., Easton, Pa.

In certain embodiments, pharmaceutical compositions suitable forparenteral administration may comprise the α5-containing GABA_(A) Rpositive allosteric modulator in combination with one or morepharmaceutically acceptable sterile isotonic aqueous or non-aqueoussolutions, dispersions, suspensions or emulsions, or sterile powderswhich may be reconstituted into sterile injectable solutions ordispersions just prior to use, which may contain antioxidants, buffers,bacteriostats, solutes which render the formulation isotonic with theblood of the intended recipient or suspending or thickening agents.Examples of suitable aqueous and non-aqueous carriers which may beemployed in the pharmaceutical compositions of the invention includewater, ethanol, polyols (such as glycerol, propylene glycol,polyethylene glycol, and the like), and suitable mixtures thereof,vegetable oils, such as olive oil, and injectable organic esters, suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of coating materials, such as lecithin, by the maintenance of therequired particle size in the case of dispersions, and by the use ofsurfactants.

A composition comprising a α5-containing GABA_(A) R positive allostericmodulator may also contain adjuvants, such as preservatives, wettingagents, emulsifying agents and dispersing agents. Prevention of theaction of microorganisms may be ensured by the inclusion of variousantibacterial and antifungal agents, for example, paraben,chlorobutanol, phenol sorbic acid, and the like. It may also bedesirable to include isotonic agents, such as sugars, sodium chloride,and the like into the compositions. In addition, prolonged absorption ofthe injectable pharmaceutical form may be brought about by the inclusionof agents which delay absorption, such as aluminum monostearate andgelatin.

In certain embodiments of the invention, compositions comprising aα5-containing GABA_(A) R positive allosteric modulator can beadministered orally, e.g., in the form of capsules, cachets, pills,tablets, lozenges (using a flavored basis, usually sucrose and acacia ortragacanth), powders, granules, or as a solution or a suspension in anaqueous or non-aqueous liquid, or as an oil-in-water or water-in-oilliquid emulsion, or as an elixir or syrup, or as pastilles (using aninert base, such as gelatin and glycerin, or sucrose and acacia) and thelike, each containing a predetermined amount of the α5-containingGABA_(A) R positive allosteric modulator as an active ingredient.

In solid dosage forms for oral administration (capsules, tablets, pills,dragees, powders, granules, and the like), one or more compositionscomprising the α5-containing GABA_(A) R positive allosteric modulatormay be mixed with one or more pharmaceutically acceptable carriers, suchas sodium citrate or dicalcium phosphate, and/or any of the following:(1) fillers or extenders, such as starches, lactose, sucrose, glucose,mannitol, and/or silicic acid; (2) binders, such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone,sucrose, and/or acacia; (3) humectants, such as glycerol; (4)disintegrating agents, such as agar-agar, calcium carbonate, potato ortapioca starch, alginic acid, certain silicates, and sodium carbonate;(5) solution retarding agents, such as paraffin; (6) absorptionaccelerators, such as quaternary ammonium compounds; (7) wetting agents,such as, for example, cetyl alcohol and glycerol monostearate; (8)absorbents, such as kaolin and bentonite clay; (9) lubricants, such atalc, calcium stearate, magnesium stearate, solid polyethylene glycols,sodium lauryl sulfate, and mixtures thereof; and (10) coloring agents.In the case of capsules, tablets and pills, the pharmaceuticalcompositions may also comprise buffering agents. Solid compositions of asimilar type may also be employed as fillers in soft and hard-filledgelatin capsules using such excipients as lactose or milk sugars, aswell as high molecular weight polyethylene glycols and the like.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups,and elixirs. In addition to the α5-containing GABA_(A) R positiveallosteric modulator, the liquid dosage forms may contain inert diluentscommonly used in the art, such as water or other solvents, solubilizingagents and emulsifiers, such as ethyl alcohol (ethanol), isopropylalcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzylbenzoate, propylene glycol, 1,3-butylene glycol, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acidesters of sorbitan, and mixtures thereof. Besides inert diluents, theoral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, coloring,perfuming, and preservative agents.

Suspensions, in addition to the active compounds, may contain suspendingagents such as ethoxylated isostearyl alcohols, polyoxyethylenesorbitol, and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, and mixturesthereof.

As described herein, the compounds, agents, and compositions thereof maybe administered for slow, controlled or extended release. The term“extended release” is widely recognized in the art of pharmaceuticalsciences and is used herein to refer to a controlled release of anactive compound or agent from a dosage form to an environment over(throughout or during) an extended period of time, e.g. greater than orequal to one hour. An extended release dosage form will release drug atsubstantially constant rate over an extended period of time or asubstantially constant amount of drug will be released incrementallyover an extended period of time. The term “extended release” used hereinincludes the terms “controlled release,” “prolonged release,” “sustainedrelease,” “delayed release,” or “slow release” as these terms are usedin the pharmaceutical sciences. In some embodiments, the extendedrelease dosage is administered in the form of a patch or a pump.

A person of ordinary skill in the art, such as a physician, is readilyable to determine the required amount of α5-containing GABA_(A) Rpositive allosteric modulator (s) to treat the subject using thecompositions and methods of the invention. It is understood that thedosage regimen will be determined for an individual, taking intoconsideration, for example, various factors that modify the action ofα5-containing GABA_(A) R positive allosteric modulator, the severity orstage of the disease, route of administration, and characteristicsunique to the individual, such as age, weight, size, and extent ofcognitive impairment.

It is well-known in the art that normalization to body surface area isan appropriate method for extrapolating doses between species. Tocalculate the human equivalent dose (HED) from a dosage used in thetreatment of age-dependent cognitive impairment in rats, the formula HED(mg/kg)=rat dose (mg/kg)×0.16 may be employed (see Estimating the SafeStarting Dose in Clinical Trials for Therapeutics in Adult HealthyVolunteers, December 2002, Center for Biologics Evaluation andResearch). For example, using that formula, a dosage of 10 mg/kg in ratsis equivalent to 1.6 mg/kg in humans. This conversion is based on a moregeneral formula HED=animal dose in mg/kg×(animal weight in kg/humanweight in kg)^(0.33).

In certain embodiments of the invention, the dose of the α5-containingGABA_(A) R positive allosteric modulator is between 0.0001 and 100mg/kg/day (which, given a typical human subject of 70 kg, is between0.007 and 7000 mg/day).

In certain embodiments of the invention, the interval of administrationis once every 12 or 24 hours. Administration at less frequent intervals,such as once every 6 hours, may also be used.

If administered by an implant, a device or a slow or extended releaseformulation, the α5-containing GABA_(A) R positive allosteric modulatorcan be administered one time, or one or more times periodicallythroughout the lifetime of the patient as necessary. Otheradministration intervals intermediate to or shorter than these dosageintervals for clinical applications may also be used and may bedetermined by one skilled in the art following the methods of thisinvention.

Desired time of administration can be determined by routineexperimentation by one skilled in the art. For example, theα5-containing GABA_(A) R positive allosteric modulator may beadministered for a period of 1-4 weeks, 1-3 months, 3-6 months, 6-12months, 1-2 years, or more, up to the lifetime of the patient.

In addition to α5-containing GABA_(A) R positive allosteric modulator,the compositions of this invention can also include othertherapeutically useful agents. These other therapeutically useful agentsmay be administered in a single formulation, simultaneously orsequentially with the α5-containing GABA_(A) R positive allostericmodulator according to the methods of the invention.

It will be understood by one of ordinary skill in the art that thecompositions described herein may be adapted and modified as isappropriate for the application being addressed and that thecompositions described herein may be employed in other suitableapplications. For example, the compositions of this application mayfurther comprise a second therapeutic agent. Such other additions andmodifications will not depart from the scope hereof.

Pharmaceutical Compositions with Antipsychotics

The compounds or the compositions of this application may be used incombination with an antipsychotic in treating cognitive impairmentassociated with schizophrenia or bipolar disorder in a subject having orat risk of said schizophrenia or bipolar disorder (e.g., mania). Theantipsychotic or a pharmaceutically acceptable salt, hydrate, solvate orpolymorph thereof that is useful in the methods and compositions of thisinvention include both typical and atypical antipsychotics. In someembodiments, the compounds or the compositions of the present inventionmay be used to treat one or more positive and/or negative symptoms, aswell as cognitive impairment, associated with schizophrenia. In someembodiments, the compounds or the compositions of the present inventionmay be used to treat one or more symptoms, as well as cognitiveimpairment, associated with bipolar disorder (in particular, mania). Insome embodiments of this invention, the compounds or the compositions ofthis invention prevent or slow the progression of cognitive impairmentof schizophrenia or bipolar disorder (in particular, mania) in saidsubject.

In some embodiments, the antipsychotics suitable for use in the presentinvention are selected from atypical antipsychotics. Such atypicalantipsychotics include, but are not limited to, those disclosed in, forexample, U.S. Pat. Nos. 4,734,416; 5,006,528; 4,145,434; 5,763,476;3,539,573; 5,229,382; 5,532,372; 4,879,288; 4,804,663; 4,710,500;4,831,031; and 5,312,925, and EP Patents EP402644 and EP368388, and thepharmaceutically acceptable salts, hydrates, solvates, and polymorphsthereof.

In some embodiments, atypical antipsychotics suitable for use in thepresent invention include, but are not limited to, aripiprazole,asenapine, clozapine, iloperidone, olanzapine, lurasidone, paliperidone,quetiapine, risperidone and ziprasidone, and the pharmaceuticallyacceptable salts, hydrates, solvates, and polymorphs thereof. In someembodiments, the antipsychotic suitable for use herein is selected fromaripiprazole (Bristol-Myers Squibb), olanzapine (Lilly) and ziprasidone(Pfizer), and the pharmaceutically acceptable salts, hydrates, solvates,and polymorphs thereof.

In some embodiments, the antipsychotics suitable for use in the presentinvention are typical antipsychotics, including, but not limited to,acepromazine, benperidol, bromazepam, bromperidol, chlorpromazine,chlorprothixene, clotiapine, cyamemazine, diazepam, dixyrazine,droperidol, flupentixol, fluphenazine, fluspirilene, haloperidol,heptaminol, isopropamide iodide, levomepromazine, levosulpiride,loxapine, melperone, mesoridazine, molindone, oxypertine, oxyprothepine,penfluridol, perazine, periciazine, perphenazine, pimozide, pipamperone,pipotiazine, prochlorperazine, promazine, promethazine, prothipendyl,pyridoxine, sulpiride, sultopride, tetrabenazine, thioproperazine,thioridazine, tiapride, tiotixene, trifluoperazine, triflupromazine,trihexyphenidyl, and zuclopenthixol, and the pharmaceutically acceptablesalts, hydrates, solvates, and polymorphs thereof.

In some embodiments of the present invention, the antipsychotic or apharmaceutically acceptable salt, hydrate, solvate or polymorph thereofmay be selected from compounds that are dopaminergic agents (such asdopamine D1 receptor antagonists or agonists, dopamine D₂ receptorantagonists or partial agonists, dopamine D3 receptor antagonists orpartial agonists, dopamine D4 receptor antagonists), glutamatergicagents, N-methyl-D-aspartate (NMDA) receptor positive allostericmodulators, glycine reuptake inhibitors, glutamate reuptake inhibitor,metabotropic glutamate receptors (mGluRs) agonists or positiveallosteric modulators (PAMs) (e.g., mGluR2/3 agonists or PAMs),glutamate receptor glur5 positive allosteric modulators (PAMs), M1muscarinic acetylcholine receptor (mAChR) positive allosteric modulators(PAMs), histamine H3 receptor antagonists,α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)/kainatereceptor antagonists, ampakines (CX-516), glutathione prodrugs,noradrenergic agents (such as alpha-2 adrenergic receptor agonists orantagonists and catechol-O-methyl transferase (COMT) inhibitors),serotonin receptor modulators (such as 5-HT_(2A) receptor antagonists,5-HT_(1A) receptor partial agonists, 5-HT_(2C) agonists, and 5-HT6antagonists, serotonin 2C agonists), cholinergic agents (such as alpha-7nicotinic receptor agonists or PAMs, alpha4-beta2 nicotinic receptoragonists, allosteric modulators of nicotinic receptors andacetylcholinesterase inhibitors, muscarinic receptor agonists andantagonists), cannabinoid CB1 antagonists, neurokinin 3 antagonists,neurotensin agonists, monoamine oxidase (MAO) B inhibitors, PDE10inhibitors, neuronal nitric oxide synthase (nNOS) inhibitors,neurosteroids, and neurotrophic factors.

In some embodiments, an α5-containing GABA_(A) receptor positiveallosteric modulator as described herein and an antipsychotic asdescribed herein, or their pharmaceutically acceptable salts, hydrates,solvates or polymorphs, are administered simultaneously, orsequentially, or in a single formulation, or in separate formulationspackaged together. In other embodiments, the α5-containing GABA_(A)receptor positive allosteric modulator and the antipsychotic, or theirpharmaceutically acceptable salts, hydrates, solvates or polymorphs, areadministered via different routes. As used herein, “combination”includes administration by any of these formulations or routes ofadministration.

Pharmaceutical Compositions with Memantine

The compounds or the compositions of this application may be used incombination with memantine or a derivative or an analog thereof intreating cognitive impairment associated with central nervous system(CNS) disorders in a subject in need or at risk thereof, including,without limitation, subjects having or at risk for age-related cognitiveimpairment, Mild Cognitive Impairment (MCI), amnestic MCI,Age-Associated Memory Impairment (AAMI), Age Related Cognitive Decline(ARCD), dementia, Alzheimer's Disease (AD), prodromal AD, post traumaticstress disorder (PTSD), schizophrenia or bipolar disorder, amyotrophiclateral sclerosis (ALS) and cancer-therapy-related cognitive impairment.

Memantine, chemically also known as 3,5-dimethyladamantan-1-amine or3,5-dimethyltricyclo[3.3.1.1^(3,7)]decan-1-amine, is an uncompetitiveN-methyl-D-aspartate (NMDA) receptor antagonist with moderate affinity.The proprietary names for memantine include: Axura® and Akatinol®(Merz), Namenda® (Forest Laboratories), Ebixa® and Abixa® (Lundbeck),and Memox® (Unipharm). Memantine is currently available in the U.S. andin over 42 countries worldwide. It is approved for the treatment ofmoderate to severe Alzheimer's disease (AD) in the United States at adose of up to 28 mg/day. Memantine and some of its derivatives andanalogs that are useful in the present invention are disclosed in U.S.Pat. Nos. 3,391,142; 4,122,193; 4,273,774; and 5,061,703, all of whichare hereby incorporated by reference. Other memantine derivatives oranalogs that are useful in the present invention include, but are notlimited to, those compounds disclosed in U.S. Patent ApplicationPublication US20040087658, US20050113458, US20060205822, US20090081259,US20090124659, and US20100227852; EP Patent Application PublicationEP2260839A2; EP Patent EP1682109B1; and PCT Application PublicationWO2005079779, all of which are incorporated herein by reference.Memantine, as used in the present invention, includes memantine and itsderivatives and analogs, as well as hydrates, polymorphs, prodrugs,salts, and solvates thereof. Memantine, as used herein, also includes acomposition comprising memantine or a derivative or an analog or apharmaceutically acceptable salt, hydrate, solvate, polymorph, orprodrug thereof, wherein the composition optionally further comprises atleast one additional therapeutic agent (such as a therapeutic agentuseful for treating a CNS disorder or cognitive impairments associatedthereof). In some embodiments, the memantine composition suitable foruse in the present invention comprises memantine and a secondtherapeutic agent that is donepezil (under the trade name Aricept).

In other embodiments of the invention, the α5-containing GABA_(A)receptor positive allosteric modulator and memantine (or the memantinederivative/analog), or their pharmaceutically acceptable salts,hydrates, solvates, polymorphs, or prodrugs are administeredsimultaneously, or sequentially, or in a single formulation or inseparate formulations packaged together. In other embodiments, theα5-containing GABA_(A) receptor positive allosteric modulator andmemantine (or the memantine derivative/analog), or theirpharmaceutically acceptable salts, hydrates, solvates, polymorphs, orprodrugs are administered via different routes. As used herein,“combination” includes administration by any of these formulations orroutes of administration.

Pharmaceutical Compositions with Acetylcholine Esterase Inhibitors(AChE-Is)

The compounds or the compositions of this application may be used incombination with an acetylcholine esterase inhibitor in treatingcognitive impairment associated with central nervous system (CNS)disorders in a subject in need or at risk thereof, including, withoutlimitation, subjects having or at risk for age-related cognitiveimpairment, Mild Cognitive Impairment (MCI), amnestic MCI,Age-Associated Memory Impairment (AAMI), Age Related Cognitive Decline(ARCD), dementia, Alzheimer's Disease (AD), prodromal AD, post traumaticstress disorder (PTSD), schizophrenia or bipolar disorder, amyotrophiclateral sclerosis (ALS) and cancer-therapy-related cognitive impairment.

AChE-Is known to a person of ordinary skill in the art may belong to thesubcategories of (i) reversible non-competitive inhibitors or reversiblecompetitive inhibitors, (ii) irreversible, and/or (iii)quasi-irreversible inhibitors.

In certain embodiment, AChE-Is useful in the present invention includethose described in PCT applications WO2014039920 and WO2002032412; EPpatents Nos. 468187; 481429-A; and U.S. Pat. Nos. 4,816,456; 4,895,841;5,041,455; 5,106,856; 5,602,176; 6,677,330; 7,340,299; 7,635,709;8,058,268; 8,741,808; and 8,853,219, all of which are incorporatedherein by reference.

In certain embodiment, typical AChE-Is that may be used in accordancewith this invention include, but are not limited to, ungeremine,ladostigil, demecarium, echothiophate (Phospholine), edrophonium(Tensilon), tacrine (Cognex), Pralidoxime (2-PAM), pyridostigmine(Mestinon), physostigmine (serine, Antilirium), abmenonium (Mytelase),galantamine (Reminyl, Razadyne), rivastigmine (Exelon, SZD-ENA-713),Huperzine A, Icopezil, neostigmine (Prostigmin, Vagostigmin), Aricept(Donepezil, E2020), Lactucopicrin, monoamine acridines and theirderivatives, piperidine and piperazine derivatives, N-benzyl-piperidinederivatives, piperidinyl-alkanoyl heterocyclic compounds,4-(1-benzyl:piperidyl)-substituted fused quinoline derivatives andcyclic amide derivatives. Other typical AChE-Is include carbamates andorganophosphonate compounds such as Metrifonate (Trichlorfon).Benzazepinols such as galantamine are also useful AChE-Is. In someembodiment, AChE-Is suitable for use in combination with the compoundsand compositions of this application include: Donepezil (aricept),Galantamine (razadyne), or Rivastigmine (exelon).

In other embodiments of the invention, the α5-containing GABA_(A)receptor positive allosteric modulator and the AChE-I, or theirpharmaceutically acceptable salts, hydrates, solvates, polymorphs, orprodrugs are administered simultaneously, or sequentially, or in asingle formulation or in separate formulations packaged together. Inother embodiments, the α5-containing GABA_(A) receptor positiveallosteric modulator and the AChE-I, or their pharmaceuticallyacceptable salts, hydrates, solvates, polymorphs, or prodrugs areadministered via different routes. As used herein, “combination”includes administration by any of these formulations or routes ofadministration.

In some embodiments, the compounds and compositions described herein arefor use as a medicament. In some embodiments, the compounds andcompositions of the present invention are for use in treating cognitiveimpairment associated with a CNS disorder in a subject in need oftreatment or at risk of said cognitive impairment. In some embodiments,the CNS disorder with cognitive impairment includes, without limitation,age-related cognitive impairment, Mild Cognitive Impairment (MCI),amnestic MCI (aMCI), Age-Associated Memory Impairment (AAMI), AgeRelated Cognitive Decline (ARCD), dementia, Alzheimer's Disease (AD),prodromal AD, post traumatic stress disorder (PTSD), schizophrenia,bipolar disorder, amyotrophic lateral sclerosis (ALS),cancer-therapy-related cognitive impairment, mental retardation,Parkinson's disease (PD), autism spectrum disorders, fragile X disorder,Rett syndrome, compulsive behavior, and substance addiction.

In some embodiments, this application provides the use of a compound orcomposition described herein in the preparation of a medicament for thetreatment of cognitive impairment associated with a CNS disorder in asubject in need of treatment or at risk of said cognitive impairment. Insome embodiments, the CNS disorder with cognitive impairment includes,without limitation, age-related cognitive impairment, Mild CognitiveImpairment (MCI), amnestic MCI (aMCI), Age-Associated Memory Impairment(AAMI), Age Related Cognitive Decline (ARCD), dementia, Alzheimer'sDisease (AD), prodromal AD, post traumatic stress disorder (PTSD),schizophrenia, bipolar disorder, amyotrophic lateral sclerosis (ALS),cancer-therapy-related cognitive impairment, mental retardation,Parkinson's disease (PD), autism spectrum disorders, fragile X disorder,Rett syndrome, compulsive behavior, and substance addiction.

Methods of Assessing Cognitive Impairment

Animal models serve as an important resource for developing andevaluating treatments for cognitive impairment associated with CNSdisorders. Features that characterize cognitive impairment in animalmodels typically extend to cognitive impairment in humans. Efficacy insuch animal models is, thus, expected to be predictive of efficacy inhumans. The extent of cognitive impairment in an animal model for a CNSdisorder, and the efficacy of a method of treatment for said CNSdisorder may be tested and confirmed with the use of a variety ofcognitive tests.

A Radial Arm Maze (RAM) behavioral task is one example of a cognitivetest, specifically testing spacial memory (Chappell et al.Neuropharmacology 37: 481-487, 1998). The RAM apparatus consists of,e.g., eight equidistantly spaced arms. A maze arm projects from eachfacet of a center platform. A food well is located at the distal end ofeach arm. Food is used as a reward. Blocks can be positioned to prevententry to any arm. Numerous extra maze cues surrounding the apparatus mayalso be provided. After habituation and training phases, spatial memoryof the subjects may be tested in the RAM under control or testcompound-treated conditions. As a part of the test, subjects arepretreated before trials with a vehicle control or one of a range ofdosages of the test compound. At the beginning of each trial, a subsetof the arms of the eight-arm maze is blocked. Subjects are allowed toobtain food on the unblocked arms to which access is permitted duringthis initial “information phase” of the trial. Subjects are then removedfrom the maze for a delay period, e.g., a 60 second delay, a 15 minutedelay, a one-hour delay, a two-hour delay, a six hour delay, a 24 hourdelay, or longer) between the information phase and the subsequent“retention test,” during which the barriers on the maze are removed,thus allowing access to all eight arms. After the delay period, subjectsare placed back onto the center platform (with the barriers to thepreviously blocked arms removed) and allowed to obtain the remainingfood rewards during this retention test phase of the trial. The identityand configuration of the blocked arms vary across trials. The number of“errors” the subjects make during the retention test phase is tracked.An error occurs in the trial if the subjects entered an arm from whichfood had already been retrieved in the pre-delay component of the trial,or if it re-visits an arm in the post-delay session that had alreadybeen visited. A fewer number of errors would indicate better spatialmemory. The number of errors made by the test subject, under varioustest compound treatment regimes, can then be compared for efficacy ofthe test compound in treating cognitive impairment associated with CNSdisorders.

Another cognitive test that may be used to assess the effects of a testcompound on the cognitive impairment of a CNS disorder model animal isthe Morris water maze. A water maze is a pool surrounded with a novelset of patterns relative to the maze. The training protocol for thewater maze may be based on a modified water maze task that has beenshown to be hippocampal-dependent (de Hoz et al., Eur. J. Neurosci.,22:745-54, 2005; Steele and Morris, Hippocampus 9:118-36, 1999). Thesubject is trained to locate a submerged escape platform hiddenunderneath the surface of the pool. During the training trial, a subjectis released in the maze (pool) from random starting positions around theperimeter of the pool. The starting position varies from trial to trial.If the subject does not locate the escape platform within a set time,the experimenter guides and places the subject on the platform to“teach” the location of the platform. After a delay period following thelast training trial, a retention test in the absence of the escapeplatform is given to assess spatial memory. The subject's level ofpreference for the location of the (now absent) escape platform, asmeasured by, e.g., the time spent in that location or the number ofcrossings of that location made by the mouse, indicates better spatialmemory, i.e., treatment of cognitive impairment. The preference for thelocation of the escape platform under different treatment conditions,can then be compared for efficacy of the test compound in treatingcognitive impairment associated with CNS disorders.

There are various tests known in the art for assessing cognitivefunction in humans, for example and without limitation, the clinicalglobal impression of change scale (CIBIC-plus scale); the Mini MentalState Exam (MMSE); the Neuropsychiatric Inventory (NPI); the ClinicalDementia Rating Scale (CDR); the Cambridge Neuropsychological TestAutomated Battery (CANTAB); the Sandoz Clinical Assessment-Geriatric(SCAG), the Buschke Selective Reminding Test (Buschke and Fuld, 1974);the Verbal Paired Associates subtest; the Logical Memory subtest; theVisual Reproduction subtest of the Wechsler Memory Scale-Revised (WMS-R)(Wechsler, 1997); the Benton Visual Retention Test, or MATRICS consensusneuropsychological test battery which includes tests of working memory,speed of processing, attention, verbal learning, visual learning,reasoning and problem solving and social cognition. See Folstein et al.,J Psychiatric Res 12: 189-98, (1975); Robbins et al., Dementia 5:266-81, (1994); Rey, L'examen clinique en psychologie, (1964); Kluger etal., J Geriatr Psychiatry Neurol 12:168-79, (1999); Marquis et al., 2002and Masur et al., 1994. Also see Buchanan, R. W., Keefe, R. S. E.,Umbricht, D., Green, M. F., Laughren, T., and Marder, S. R. (2011) TheFDA-NIMH-MATRICS guidelines for clinical trial design ofcognitive-enhancing drugs: what do we know 5 years later? Schizophr.Bull. 37, 1209-1217. Another example of a cognitive test in humans isthe explicit 3-alternative forced choice task. In this test, subjectsare presented with color photographs of common objects consisting of amix of three types of image pairs: similar pairs, identical pairs andunrelated foils. The second of the pair of similar objects is referredto as the “lure”. These image pairs are fully randomized and presentedindividually as a series of images. Subjects are instructed to make ajudgment as to whether the objects seen are new, old or similar. A“similar” response to the presentation of a lure stimulus indicatessuccessful memory retrieval by the subject. By contrast, calling thelure stimulus “old” or “new” indicates that correct memory retrieval didnot occur.

In addition to assessing cognitive performance, the progression ofage-related cognitive impairment and dementia, as well as the conversionof age-related cognitive impairment into dementia, may be monitored byassessing surrogate changes in the brain of the subject. Surrogatechanges include, without limitation, changes in regional brain volumes,perforant path degradation, and changes seen in brain function throughresting state fMRI (R-fMRI) and fluorodeoxyglucose positron emissiontomography (FDG-PET). Examples of regional brain volumes useful inmonitoring the progression of age-related cognitive impairment anddementia include reduction of hippocampal volume and reduction in volumeor thickness of entorhinal cortex. These volumes may be measured in asubject by, for example, MRI. Aisen et al., Alzheimer's & Dementia6:239-246 (2010). Perforant path degradation has been shown to be linkedto age, as well as reduced cognitive function. For example, older adultswith more perforant path degradation tend to perform worse inhippocampus-dependent memory tests. Perforant path degradation may bemonitored in subjects through ultrahigh-resolution diffusion tensorimaging (DTI). Yassa et al., PNAS 107:12687-12691 (2010). Resting-statefMRI (R-fMRI) involves imaging the brain during rest, and recordinglarge-amplitude spontaneous low-frequency (<0.1 Hz) fluctuations in thefMRI signal that are temporally correlated across functionally relatedareas. Seed-based functional connectivity, independent componentanalyses, and/or frequency-domain analyses of the signals are used toreveal functional connectivity between brain areas, particularly thoseareas whose connectivity increase or decrease with age, as well as theextent of cognitive impairment and/or dementia. FDG-PET uses the uptakeof FDG as a measure of regional metabolic activity in the brain. Declineof FDG uptake in regions such as the posterior cingulated cortex,temporoparietal cortex, and prefrontal association cortex has been shownto relate to the extent of cognitive decline and dementia. Aisen et al.,Alzheimer's & Dementia 6:239-246 (2010), Herholz et al., NeuroImage17:302-316 (2002).

Age-Related Cognitive Impairment

The invention provides methods and compositions for treating age-relatedcognitive impairment or the risk thereof using a α5-containing GABA_(A)receptor positive allosteric modulator (i.e., a compound of theinvention), such as one selected from the compounds or pharmaceuticallyacceptable salts, hydrates, solvates, polymorphs, isomers, orcombinations thereof as described herein. In certain embodiments,treatment comprises preventing or slowing the progression, ofage-related cognitive impairment. In certain embodiments, treatmentcomprises alleviation, amelioration or slowing the progression, of oneor more symptoms associated with age-related cognitive impairment. Incertain embodiments, treatment of age-related cognitive impairmentcomprises slowing the conversion of age-related cognitive impairment(including, but not limited to MCI, ARCD and AAMI) into dementia (e.g.,AD). The methods and compositions may be used for human patients inclinical applications in the treating age-related cognitive impairmentin conditions such as MCI, ARCD and AAMI or for the risk thereof. Thedose of the composition and dosage interval for the method is, asdescribed herein, one that is safe and efficacious in thoseapplications. In some embodiments of the invention, there is provided amethod of preserving or improving cognitive function in a subject withage-related cognitive impairment, the method comprising the step ofadministering to said subject a therapeutically effective amount of acompound of the invention or a pharmaceutically acceptable salt,hydrate, solvate, polymorph, isomer, or combination thereof.

In some embodiments, a subject to be treated by the methods andcompositions of this invention exhibits age-related cognitive impairmentor is at risk of such impairment. In some embodiments, the age-relatedcognitive impairment includes, without limitation, Age-Associated MemoryImpairment (AAMI), Mild Cognitive Impairment (MCI) and Age-relatedCognitive Decline (ARCD).

Animal models serve as an important resource for developing andevaluating treatments for such age-related cognitive impairments.Features that characterize age-related cognitive impairment in animalmodels typically extend to age-related cognitive impairment in humans.Efficacy in such animal models is, thus, expected to be predictive ofefficacy in humans.

Various animal models of age-related cognitive impairment are known inthe art. For example, extensive behavioral characterization hasidentified a naturally occurring form of cognitive impairment in anoutbred strain of aged Long-Evans rats (Charles River Laboratories;Gallagher et al., Behav. Neurosci. 107:618-626, (1993)). In a behavioralassessment with the Morris Water Maze (MWM), rats learn and remember thelocation of an escape platform guided by a configuration of spatial cuessurrounding the maze. The cognitive basis of performance is tested inprobe trials using measures of the animal's spatial bias in searchingfor the location of the escape platform. Aged rats in the studypopulation have no difficulty swimming to a visible platform, but anage-dependent impairment is detected when the platform is camouflaged,requiring the use of spatial information. Performance for individualaged rats in the outbred Long-Evans strain varies greatly. For example,a proportion of those rats perform on a par with young adults. However,approximately 40-50% fall outside the range of young performance. Thisvariability among aged rats reflects reliable individual differences.Thus, within the aged population some animals are cognitively impairedand designated aged-impaired (AI) and other animals are not impaired andare designated aged-unimpaired (AU). See, e.g., Colombo et al., Proc.Natl. Acad. Sci. 94: 14195-14199, (1997); Gallagher and Burwell,Neurobiol. Aging 10: 691-708, (1989); Gallagher et al. Behav. Neurosci.107:618-626, (1993); Rapp and Gallagher, Proc. Natl. Acad. Sci. 93:9926-9930, (1996); Nicolle et al., Neuroscience 74: 741-756, (1996);Nicolle et al., J. Neurosci. 19: 9604-9610, (1999); International PatentPublication WO2007/019312 and International Patent Publication WO2004/048551. Such an animal model of age-related cognitive impairmentmay be used to assay the effectiveness of the methods and compositionsthis invention in treating age-related cognitive impairment.

The efficacy of the methods and compositions of this invention intreating age-related cognitive impairment may be assessed using avariety of cognitive tests, including the Morris water maze and theradial arm maze, as discussed herein.

Dementia

The invention also provides methods and compositions for treatingdementia using a α5-containing GABA_(A) receptor positive allostericmodulator, such as one selected from the compounds or pharmaceuticallyacceptable salts, hydrates, solvates, polymorphs, isomers, orcombinations thereof as described herein. In certain embodiments,treatment comprises preventing or slowing the progression, of dementia.In certain embodiments, treatment comprises alleviation, amelioration,or slowing the progression of one or more symptoms associated withdementia. In certain embodiments, the symptom to be treated is cognitiveimpairment. In some embodiments of the invention, there is provided amethod of preserving or improving cognitive function in a subject withdementia, the method comprising the step of administering to saidsubject a therapeutically effective amount of a compound of theinvention or a pharmaceutically acceptable salt, hydrate, solvate,polymorph, isomer, or combination thereof. In certain embodiments, thedementia is Alzheimer's disease (AD), vascular dementia, dementia withLewy bodies, or frontotemporal dementia. The methods and compositionsmay be used for human patients in clinical applications in treatingdementia. The dose of the composition and dosage interval for the methodis, as described herein, one that is safe and efficacious in thoseapplications.

Animal models serve as an important resource for developing andevaluating treatments for dementia. Features that characterize dementiain animal models typically extend to dementia in humans. Thus, efficacyin such animal models is expected to be predictive of efficacy inhumans. Various animal models of dementia are known in the art, such asthe PDAPP, Tg2576, APP23, TgCRND8, J20, hPS2 Tg, and APP+PS1 transgenicmice. Sankaranarayanan, Curr. Top. Medicinal Chem. 6: 609-627, 2006;Kobayashi et al. Genes Brain Behav. 4: 173-196. 2005; Ashe and Zahns,Neuron. 66: 631-45, 2010. Such animal models of dementia may be used toassay the effectiveness of the methods and compositions of thisinvention of the invention in treating dementia.

The efficacy of the methods and compositions of this invention intreating dementia, or cognitive impairment associated with dementia, maybe assessed in animals models of dementia, as well as human subjectswith dementia, using a variety of cognitive tests known in the art, asdiscussed herein.

Post Traumatic Stress Disorder

The invention also provides methods and compositions for treating posttraumatic stress disorder (PTSD) using a α5-containing GABA_(A) receptorpositive allosteric modulator, such as one selected from the compoundsor pharmaceutically acceptable salts, hydrates, solvates, polymorphs,isomers, or combinations thereof as described herein. In certainembodiments, treatment comprises preventing or slowing the progression,of PTSD. In certain embodiments, treatment comprises alleviation,amelioration, or slowing the progression of one or more symptomsassociated with PTSD. In certain embodiments, the symptom to be treatedis cognitive impairment. In some embodiments of the invention, there isprovided a method of preserving or improving cognitive function in asubject with PTSD, the method comprising the step of administering tosaid subject a therapeutically effective amount of a compound of theinvention or a pharmaceutically acceptable salt, hydrate, solvate,polymorph, isomer, or combination thereof. The methods and compositionsmay be used for human patients in clinical applications in treatingPTSD. The dose of the composition and dosage interval for the method is,as described herein, one that is safe and efficacious in thoseapplications.

Patients with PTSD (and, to a lesser degree trauma-exposed patientswithout PTSD) have smaller hippocampal volumes (Woon et al., Prog.Neuro-Psychopharm. & Biological Psych. 34, 1181-1188; Wang et al., Arch.Gen. Psychiatry 67:296-303, 2010). PTSD is also associated with impairedcognitive performance. Older individuals with PTSD have greater declinesin cognitive performance relative to control patients (Yehuda et al.,Bio. Psych. 60: 714-721, 2006) and have a greater likelihood ofdeveloping dementia (Yaffe et al., Arch. Gen. Psych. 678: 608-613,2010).

Animal models serve as an important resource for developing andevaluating treatments for PTSD. Features that characterize PTSD inanimal models typically extend to PTSD in humans. Thus, efficacy in suchanimal models is expected to be predictive of efficacy in humans.Various animal models of PTSD are known in the art.

One rat model of PTSD is Time-dependent sensitization (TDS). TDSinvolves exposure of the animal to a severely stressful event followedby a situational reminder of the prior stress. The following is anexample of TDS. Rats are placed in a restrainer, then placed in a swimtank and made to swim for a period of time, e.g., 20 min. Followingthis, each rat is then immediately exposed to a gaseous anesthetic untilloss of consciousness, and finally dried. The animals are leftundisturbed for a number of days, e.g., one week. The rats are thenexposed to a “restress” session consisting of an initial stressor, e.g.,a swimming session in the swim tank (Liberzon et al.,Psychoneuroendocrinology 22: 443-453, 1997; Harvery et al.,Psychopharmacology 175:494-502, 2004). TDS results in an enhancement ofthe acoustic startle response (ASR) in the rat, which is comparable tothe exaggerated acoustic startle that is a prominent symptom of PTSD(Khan and Liberzon, Psychopharmacology 172: 225-229, 2004). Such animalmodels of PTSD may be used to assay the effectiveness of the methods andcompositions of this invention of the invention in treating PTSD.

The efficacy of the methods and compositions of this invention intreating PTSD, or cognitive impairment associated with PTSD, may also beassessed in animals models of PTSD, as well as human subjects with PTSD,using a variety of cognitive tests known in the art, as discussedherein.

Schizophrenia and Bipolar Disorder

The invention additionally provides methods and compositions fortreating schizophrenia or bipolar disorder (in particular, mania) usinga α5-containing GABA_(A) receptor positive allosteric modulator, such asone selected from the compounds or pharmaceutically acceptable salts,hydrates, solvates, polymorphs, isomers, or combinations thereof asdescribed herein. In certain embodiments, treatment comprises preventingor slowing the progression of schizophrenia or bipolar disorder (inparticular, mania). Schizophrenia is characterized by a wide spectrum ofpsychopathology, including positive symptoms such as aberrant ordistorted mental representations (e.g., hallucinations, delusions), ordopamine dysregulation-associated symptoms (e.g., hyperdopaminergicresponses, hyperdopaminergic behavioral responses, dopaminergichyperactivity, or hyperlocomotor activity, or psychosis), negativesymptoms characterized by diminution of motivation and adaptivegoal-directed action (e.g., anhedonia, affective flattening, avolition),and cognitive impairment. In certain embodiments, treatment comprisesalleviation, amelioration or slowing the progression of one or morepositive and/or negative symptoms, as well as cognitive impairment,associated with schizophrenia. Further, there are a number of otherpsychiatric diseases such as schizotypical and schizoaffective disorder,other acute- and chronic psychoses and bipolar disorder (in particular,mania), which have an overlapping symptomatology with schizophrenia. Insome embodiments, treatment comprises alleviation, amelioration orslowing the progression of one or more symptoms, as well as cognitiveimpairment, associated with bipolar disorder (in particular, mania). Insome embodiments of the invention, there is provided a method ofpreserving or improving cognitive function in a subject withschizophrenia or bipolar disorder, the method comprising the step ofadministering to said subject a therapeutically effective amount of acompound of the invention or a pharmaceutically acceptable salt,hydrate, solvate, polymorph, isomer, or combination thereof. The methodsand compositions may be used for human patients in clinical applicationsin treating schizophrenia or bipolar disorder (in particular, mania).The dose of the composition and dosage interval for the method is, asdescribed herein, one that is safe and efficacious in thoseapplications.

Cognitive impairments are associated with schizophrenia. They precedethe onset of psychosis and are present in non-affected relatives. Thecognitive impairments associated with schizophrenia constitute a goodpredictor for functional outcome and are a core feature of the disorder.Cognitive features in schizophrenia reflect dysfunction in frontalcortical and hippocampal circuits. Patients with schizophrenia alsopresent hippocampal pathologies such as reductions in hippocampalvolume, reductions in neuronal size and dysfunctional hyperactivity. Animbalance in excitation and inhibition in these brain regions has alsobeen documented in schizophrenic patients suggesting that drugstargeting inhibitory mechanisms could be therapeutic. See, e.g.,Guidotti et al., Psychopharmacology 180: 191-205, 2005; Zierhut, Psych.Res. Neuroimag. 183:187-194, 2010; Wood et al., NeuroImage 52:62-63,2010; Vinkers et al., Expert Opin. Investig. Drugs 19:1217-1233, 2009;Young et al., Pharmacol. Ther. 122:150-202, 2009.

Animal models serve as an important resource for developing andevaluating treatments for schizophrenia. Features that characterizeschizophrenia in animal models typically extend to schizophrenia inhumans. Thus, efficacy in such animal models is expected to bepredictive of efficacy in humans. Various animal models of schizophreniaare known in the art.

One animal model of schizophrenia is protracted treatment withmethionine. Methionine-treated mice exhibit deficient expression ofGAD67 in frontal cortex and hippocampus, similar to those reported inthe brain of postmortem schizophrenia patients. They also exhibitprepulse inhibition of startle and social interaction deficits(Tremonlizzo et al., PNAS, 99: 17095-17100, 2002). Another animal modelof schizophrenia is methylaoxymethanol acetate (MAM)-treatment in rats.Pregnant female rats are administered MAM (20 mg/kg, intraperitoneal) ongestational day 17. MAM-treatment recapitulate a pathodevelopmentalprocess to schizophrenia-like phenotypes in the offspring, includinganatomical changes, behavioral deficits and altered neuronal informationprocessing. More specifically, MAM-treated rats display a decreaseddensity of parvalbumin-positive GABAergic interneurons in portions ofthe prefrontal cortex and hippocampus. In behavioral tests, MAM-treatedrats display reduced latent inhibition. Latent inhibition is abehavioral phenomenon where there is reduced learning about a stimulusto which there has been prior exposure with any consequence. Thistendency to disregard previously benign stimuli, and reduce theformation of association with such stimuli is believed to preventsensory overload. Low latent inhibition is indicative of psychosis.Latent inhibition may be tested in rats in the following manner. Ratsare divided into two groups. One group is pre-exposed to a tone overmultiple trials. The other group has no tone presentation. Both groupsare then exposed to an auditory fear conditioning procedure, in whichthe same tone is presented concurrently with a noxious stimulus, e.g. anelectric shock to the foot. Subsequently, both groups are presented withthe tone, and the rats' change in locomotor activity during tonepresentation is monitored. After the fear conditioning the rats respondto the tone presentation by strongly reducing locomotor activity.However, the group that has been exposed to the tone before theconditioning period displays robust latent inhibition: the suppressionof locomotor activity in response to tone presentation is reduced.MAM-treated rats, by contrast show impaired latent inhibition. That is,exposure to the tone previous to the fear conditioning procedure has nosignificant effect in suppressing the fear conditioning. (see Lodge etal., J. Neurosci., 29:2344-2354, 2009) Such animal models ofschizophrenia may be used to assay the effectiveness of the methods andcompositions of the invention in treating schizophrenia or bipolardisorder (in particular, mania).

MAM-treated rats display a significantly enhanced locomotor response (oraberrant locomotor activity) to low dose D-amphetamine administration.The MAM-treated rats also display a significantly greater number ofspontaneously firing ventral tegmental dopamine (DA) neurons. Theseresults are believed to be a consequence of excessive hippocampalactivity because in MAM-treated rats, the ventral hippocampus (vHipp)inactivation (e.g., by intra-vHipp administration of a sodium channelblocker, tetrodotoxin (TTX), to MAM rats) completely reversed theelevated DA neuron population activity and also normalized the augmentedamphetamine-induced locomotor behavior. The correlation of hippocampaldysfunction and the hyper-responsivity of the DA system is believed tounderlie the augmented response to amphetamine in MAM-treated animalsand psychosis in schizophrenia patients. See Lodge D. J. et al.Neurobiology of Disease (2007), 27(42), 11424-11430. The use ofMAM-treated rats in the above study may be suitable for use to assay theeffectiveness of the methods and compositions of the present inventionin treating schizophrenia or bipolar disorder (in particular, mania).For example, the methods and compositions of this invention maybeevaluated, using MAM-treated animals, for their effects on the centralhippocampus (vHipp) regulation, on the elevated DA neuron populationactivity and on the hyperactive locomotor response to amphetamine in theMAM-treated animals.

In MAM-treated rats, hippocampal (HPC) dysfunction leads to dopaminesystem hyperactivity. A benzodiazepine-positive allosteric modulator(PAM), selective for the α5 subunit of the GABA_(A) receptor,SH-053-2′F—R—CH₃, is tested for its effects on the output of thehippocampal (HPC). The effect of SH-053-2′F—R—CH₃ on the hyperactivelocomotor response to amphetamine in MAM-treated animals is alsoexamined. The α5GABAAR PAM reduces the number of spontaneously active DAneurons in the ventral tegmental area (VTA) of MAM rats to levelsobserved in saline-treated rats (control group), both when administeredsystemically and when directly infused into the ventral HPC. Moreover,HPC neurons in both saline-treated and MAM-treated animals showdiminished cortical-evoked responses following the α5GABAAR PAMtreatment. In addition, the increased locomotor response to amphetamineobserved in MAM-treated rats is reduced following the α5GABA_(A)R PAMtreatment. See Gill K. M et al. Neuropsychopharmacology (2011), 1-9. Theuse of MAM-treated rats in the above study may be suitable for use inthe present invention to assay the effectiveness of the methods andcompositions of the invention in treating schizophrenia or bipolardisorder (in particular, mania). For example, the methods andcompositions of this invention maybe evaluated, using MAM-treatedanimals, for their effects on the output of the hippocampal (HPC) and onthe hyperactive locomotor response to amphetamine in the MAM-treatedanimals.

Administration of MAM to pregnant rats on embryonic day 15 (E15)severely impairs spatial memory or the ability to learn the spatiallocation of four items on an eight-arm radial maze in the offspring. Inaddition, embryonic day 17 (E17) MAM-treated rats are able to reach thelevel of performance of control rats at the initial stages of training,but are unable to process and retrieve spatial information when a 30-mindelay is interposed, indicating a significant impairment in workingmemory. See Gourevitch R. et al. (2004). Behav. Pharmacol, 15, 287-292.Such animal models of schizophrenia may be used to assay theeffectiveness of the methods and compositions of the invention intreating schizophrenia or bipolar disorder (in particular, mania).

Apomorphine-induced climbing (AIC) and stereotype (AIS) in mice isanother animal model useful in this invention. Agents are administeredto mice at a desired dose level (e.g., via intraperitonealadministration). Subsequently, e.g., thirty minutes later, experimentalmice are challenges with apomorphine (e.g., with 1 mg/kg sc). Fiveminutes after the apomorphine injection, the sniffing-licking-gnawingsyndrome (stereotyped behavior) and climbing behavior induced byapomorphine are scored and recorded for each animal. Readings can berepeated every 5 min during a 30-min test session. Scores for eachanimal are totaled over the 30-min test session for each syndrome(stereotyped behavior and climbing). If an effect reached at least of50% inhibition, and ID₅₀ value (95% confidence interval) is calculatedusing a nonlinear least squares calculation with inverse prediction.Mean climbing and stereotype scores can be expressed as a percent ofcontrol values observed in vehicle treated (e.g., saline-treated) micethat receive apomorphine. See Grauer S. M. et al. Psychopharmacology(2009) 204, 37-48. This mouse model may be used to assay theeffectiveness of the methods and compositions of the invention intreating schizophrenia or bipolar disorder (in particular, mania).

In another well-established preclinical model of schizophrenia, ratsexposed chronically to ketamine, an uncompetitive N-methyl-D-aspartate(NMDA) receptor antagonist, produces positive and negative psychoticsymptoms and cognitive impairment. Long-Evans male rats are injectedintraperitoneally with ketamine (30 mg/kg, twice a day) for two weeksduring adolescence (2 month-old). Rats are behaviorally tested when theyreach adulthood (approximately 4-5 month-old) for the behavioralsymptoms to ketamine exposure and for the efficacy of treatment toalleviate those symptoms. See, e.g., Enomoto et al. Progress inNeuro-Psychopharmacology & Biological Psychiatry 33 (2009) 668-675.

The efficacy of the methods and compositions of this invention intreating schizophrenia or cognitive impairment associated therewith mayalso be assessed in animal models of schizophrenia or bipolar disorder(in particular, mania), as well as human subjects with schizophrenia,using a variety of cognitive tests known in the art, as discussedherein.

Amyotrophic Lateral Sclerosis (ALS)

The invention additionally provides methods and compositions fortreating ALS using a α5-containing GABA_(A) receptor positive allostericmodulator, such as one selected from the compounds or pharmaceuticallyacceptable salts, hydrates, solvates, polymorphs, isomers, orcombinations thereof as described herein. In certain embodiments,treatment comprises preventing or slowing the progression, of ALS. Incertain embodiments, treatment comprises alleviation, amelioration orslowing the progression, of one or more symptoms associated with ALS. Incertain embodiments, the symptom to be treated is cognitive impairment.In some embodiments of the invention, there is provided a method ofpreserving or improving cognitive function in a subject with ALS, themethod comprising the step of administering to said subject atherapeutically effective amount of a compound of the invention or apharmaceutically acceptable salt, hydrate, solvate, polymorph, isomer,or combination thereof. The methods and compositions may be used forhuman patients in clinical applications in treating ALS. The dose of thecomposition and dosage interval for the method is, as described herein,one that is safe and efficacious in those applications.

In addition to the degeneration of motor neurons, ALS is characterizedby neuronal degeneration in the entorhinal cortex and hippocampus,memory deficits, and neuronal hyperexcitability in different brain areassuch as the cortex.

The efficacy of the methods and compositions of this invention intreating ALS, or cognitive impairment associated with ALS, may also beassessed in animal models of ALS, as well as human subjects with ALS,using a variety of cognitive tests known in the art, as discussedherein.

Cancer Therapy-Related Cognitive Impairment

The invention additionally provides methods and compositions fortreating cancer therapy-related cognitive impairment using aα5-containing GABA_(A) receptor positive allosteric modulator, such asone selected from the compounds or pharmaceutically acceptable salts,hydrates, solvates, polymorphs, isomers, or combinations thereof asdescribed herein. In certain embodiments, treatment comprises preventingor slowing the progression, of cancer therapy-related cognitiveimpairment. In certain embodiments, treatment comprises alleviation,amelioration or slowing the progression, of one or more symptomsassociated with cancer therapy-related cognitive impairment. In someembodiments of the invention, there is provided a method of preservingor improving cognitive function in a subject with cancer therapy-relatedcognitive impairment, the method comprising the step of administering tosaid subject a therapeutically effective amount of a compound of theinvention or a pharmaceutically acceptable salt, hydrate, solvate,polymorph, isomer, or combination thereof. The methods and compositionsmay be used for human patients in clinical applications in treatingcancer therapy-related cognitive impairment. The dose of the compositionand dosage interval for the method is, as described herein, one that issafe and efficacious in those applications.

Therapies that are used in cancer treatment, including chemotherapy,radiation, or combinations thereof, can cause cognitive impairment inpatients, in such functions as memory, learning and attention.Cytotoxicity and other adverse side-effects on the brain of cancertherapies are the basis for this form of cognitive impairment, which canpersist for decades. (Dietrich et al., Oncologist 13:1285-95, 2008;Soussain et al., Lancet 374:1639-51, 2009).

Cognitive impairment following cancer therapies reflects dysfunction infrontal cortical and hippocampal circuits that are essential for normalcognition. In animal models, exposure to either chemotherapy orradiation adversely affects performance on tests of cognitionspecifically dependent on these brain systems, especially thehippocampus (Kim et al., J. Radiat. Res. 49:517-526, 2008; Yang et al.,Neurobiol. Learning and Mem. 93:487-494, 2010). Thus, drugs targetingthese cortical and hippocampal systems could be neuroprotective inpatients receiving cancer therapies and efficacious in treating symptomsof cognitive impairment that may last beyond the interventions used ascancer therapies.

Animal models serve as an important resource for developing andevaluating treatments for cancer therapy-related cognitive impairment.Features that characterize cancer therapy-related cognitive impairmentin animal models typically extend to cancer therapy-related cognitiveimpairment in humans. Thus, efficacy in such animal models is expectedto be predictive of efficacy in humans. Various animal models of cancertherapy-related cognitive impairment are known in the art.

Examples of animal models of cancer therapy-related cognitive impairmentinclude treating animals with anti-neoplastic agents such ascyclophosphamide (CYP) or with radiation, e.g., ⁶⁰Co gamma-rays. (Kim etal., J. Radiat. Res. 49:517-526, 2008; Yang et al., Neurobiol. Learningand Mem. 93:487-494, 2010). The cognitive function of animal models ofcancer therapy-related cognitive impairment may then be tested withcognitive tests to assay the effectiveness of the methods andcompositions of the invention in treating cancer therapy-relatedcognitive impairment. The efficacy of the methods and compositions ofthis invention in treating cancer therapy-related cognitive impairment,as well as human subjects with cancer therapy-related cognitiveimpairment, using a variety of cognitive tests known in the art, asdiscussed herein.

Parkinson's Disease (PD)

Parkinson's disease (PD) is a neurological disorder characterized by adecrease of voluntary movements. The afflicted patient has reduction ofmotor activity and slower voluntary movements compared to the normalindividual. The patient has characteristic “mask” face, a tendency tohurry while walking, bent over posture and generalized weakness of themuscles. There is a typical “lead-pipe” rigidity of passive movements.Another important feature of the disease is the tremor of theextremities occurring at rest and decreasing during movements.

Parkinson's disease, the etiology of which is unknown, belongs to agroup of the most common movement disorders named parkinsonism, whichaffects approximately one person per one thousand. These other disordersgrouped under the name of parkinsonism may result from viral infection,syphilis, arteriosclerosis and trauma and exposure to toxic chemicalsand narcotics. Nonetheless, it is believed that the inappropriate lossof synaptic stability may lead to the disruption of neuronal circuitsand to brain diseases. Whether as the result of genetics, drug use, theaging process, viral infections, or other various causes, dysfunction inneuronal communication is considered the underlying cause for manyneurologic diseases, such as PD (Myrrhe van Spronsen and Casper C.Hoogenraad, Curr. Neurol. Neurosci. Rep. 2010, 10, 207-214).

Regardless of the cause of the disease, the main pathologic feature isdegeneration of dopaminergic cells in basal ganglia, especially insubstantia nigra. Due to premature death of the dopamine containingneurons in substantia nigra, the largest structure of the basal ganglia,the striatum, will have reduced input from substantia nigra resulting indecreased dopamine release. The understanding of the underlyingpathology led to the introduction of the first successful treatmentwhich can alleviate Parkinson's disease. Virtually all approaches to thetherapy of the disease are based on dopamine replacement. Drugscurrently used in the treatment can be converted into dopamine aftercrossing the blood brain barrier, or they can boost the synthesis ofdopamine and reduce its breakdown. Unfortunately, the main pathologicevent, degeneration of the cells in substantia nigra, is not helped. Thedisease continues to progress and frequently after a certain length oftime, dopamine replacement treatment will lose its effectiveness.

The invention provides methods and compositions for treating PD using aα5-containing GABA_(A) receptor positive allosteric modulator, such asone selected from the compounds or pharmaceutically acceptable salts,hydrates, solvates, polymorphs, isomers, or combinations thereof asdescribed herein. In certain embodiments, treatment comprises preventingor slowing the progression of PD. In certain embodiments, treatmentcomprises alleviation, amelioration, or slowing the progression of oneor more symptoms associated with PD. In certain embodiments, the symptomto be treated is cognitive impairment. For example, methods andcompositions of the disclosure can be used to improve themotor/cognitive impairments symptomatic of Parkinson's disease.Moreover, methods and compositions of the disclosure may be useful fortreating the memory impairment symptomatic of Parkinson's disease. Insome embodiments of the invention, there is provided a method ofpreserving or improving cognitive function in a subject with PD, themethod comprising the step of administering to said subject atherapeutically effective amount of a compound of the invention or apharmaceutically acceptable salt, hydrate, solvate, polymorph, isomer,or combination thereof.

There are a number of animal models for PD. Exemplary animal models forPD include the reserpine model, the methamphetamine model, the6-hydroxydopamine (6-OHDA) model, the1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) model, the paraquat(PQ)-Maneb model, the rotenone model, the 3-nitrotyrosine model andgenetic models using transgenic mice. Transgenic models include micethat over express α-synuclein, express human mutant forms ofα-synuclein, or mice that express LRKK2 mutations. See review of thesemodels by Ranjita B. et al. (Ranjita B. et al. BioEssays 2002, 24,308-318). Additional information regarding these animal models isreadily available from Jackson Laboratories (see alsohttp://research.jax.org/grs/parkinsons.html), as well as in numerouspublications disclosing the use of these validated models.

The efficacy of the methods and compositions of this invention intreating PD, or cognitive impairment associated with PD, may be assessedin any of the above animal models of PD, as well as human subjects withPD, using a variety of cognitive tests known in the art, as discussedherein.

Autism

Autism is a neurodevelopmental disorder characterized by dysfunction inthree core behavioral dimensions: repetitive behaviors, social deficits,and cognitive deficits. The repetitive behavior domain involvescompulsive behaviors, unusual attachments to objects, rigid adherence toroutines or rituals, and repetitive motor mannerisms such asstereotypies and self-stimulatory behaviors. The social deficitdimension involves deficits in reciprocal social interactions, lack ofeye contact, diminished ability to carry on conversation, and impaireddaily interaction skills. The cognitive deficits can include languageabnormalities. Autism is a disabling neurological disorder that affectsthousands of Americans and encompasses a number of subtypes, withvarious putative causes and few documented ameliorative treatments. Thedisorders of the autistic spectrum may be present at birth, or may havelater onset, for example, at ages two or three. There are no clear cutbiological markers for autism. Diagnosis of the disorder is made byconsidering the degree to which the child matches the behavioralsyndrome, which is characterized by poor communicative abilities,peculiarities in social and cognitive capacities, and maladaptivebehavioral patterns. The dysfunction in neuronal communication isconsidered one of the underlying causes for autism (Myrrhe van Spronsenand Casper C. Hoogenraad, Curr. Neurol. Neurosci. Rep. 2010, 10,207-214). Recent studies have shown that there is a GABA_(A) α5 deficitin autism spectrum disorder (ASD) and support further investigations ofthe GABA system in this disorder (Mendez M A, et al. Neuropharmacology.2013, 68:195-201).

The invention also provides methods and compositions for treating autismusing a α5-containing GABA_(A) receptor positive allosteric modulator,such as one selected from the compounds or pharmaceutically acceptablesalts, hydrates, solvates, polymorphs, isomers, or combinations thereofas described herein. In certain embodiments, treatment comprisespreventing or slowing the progression of autism. In certain embodiments,treatment comprises alleviation, amelioration, or slowing theprogression of one or more symptoms associated with autism. In certainembodiments, the symptom to be treated is cognitive impairment orcognitive deficit. For example, methods and compositions of thedisclosure can be used to improve the motor/cognitive deficitssymptomatic of autism. In some embodiments of the invention, there isprovided a method of preserving or improving cognitive function in asubject with autism, the method comprising the step of administering tosaid subject a therapeutically effective amount of a compound of theinvention or a pharmaceutically acceptable salt, hydrate, solvate,polymorph, isomer, or combination thereof.

The valproic acid (VPA) rat model of autism using in vitroelectrophysiological techniques, established by Rodier et al. (Rodier,P. M. et al. Reprod. Toxicol. 1997, 11, 417-422) is one of the mostexhaustively established insult-based animal models of autism and isbased on the observation that pregnant women treated with VPA in the1960s, during a circumscribed time window of embryogenesis, had a muchhigher risk of giving birth to an autistic child than the normalpopulation. Offspring of VPA-exposed pregnant rats show severalanatomical and behavioral symptoms typical of autism, such as diminishednumber of cerebellar Purkinje neurons, impaired social interaction,repetitive behaviors as well as other symptoms of autism, includingenhanced fear memory processing. See, Rinaldi T. et al. Frontiers inNeural Circuits, 2008, 2, 1-7. Another mouse model, BTBR T+tf/J (BTBR)mice, an established model with robust behavioral phenotypes relevant tothe three diagnostic behavioral symptoms of autism—unusual socialinteractions, impaired communication, and repetitive behaviors—was usedto probe the efficacy of a selective negative allosteric modulator ofthe mGluR5 receptor, GRN-529. See, e.g., Silverman J. L. et al. SciTransl. Med. 2012, 4, 131. The efficacy of the methods and compositionsof this invention in treating autism, or cognitive deficits associatedwith autism, may be assessed in the VPA-treated rat model of autism orthe BTBR T+tf/J (BTBR) mouse model, as well as human subjects withautism, using a variety of cognitive tests known in the art, asdiscussed herein.

Mental Retardation

Mental retardation is a generalized disorder characterized bysignificantly impaired cognitive function and deficits in adaptivebehaviors. Mental retardation is often defined as an IntelligenceQuotient (IQ) score of less than 70. Inborn causes are among manyunderlying causes for mental retardation. The dysfunction in neuronalcommunication is also considered one of the underlying causes for mentalretardation (Myrrhe van Spronsen and Casper C. Hoogenraad, Curr. Neurol.Neurosci. Rep. 2010, 10, 207-214).

In some instances, mental retardation includes, but are not limited to,Down syndrome, velocariofacial syndrome, fetal alcohol syndrome, FragileX syndrome, Klinefelter's syndrome, neurofibromatosis, congenitalhypothyroidism, Williams syndrome, phenylketonuria (PKU),Smith-Lemli-Opitz syndrome, Prader-Willi syndrome, Phelan-McDermidsyndrome, Mowat-Wilson syndrome, ciliopathy, Lowe syndrome and sideriumtype X-linked mental retardation. Down syndrome is a disorder thatincludes a combination of birth defects, including some degree of mentalretardation, characteristic facial features and, often, heart defects,increased infections, problems with vision and hearing, and other healthproblems. Fragile X syndrome is a prevalent form of inherited mentalretardation, occurring with a frequency of 1 in 4,000 males and 1 in8,000 females. The syndrome is also characterized by developmentaldelay, hyperactivity, attention deficit disorder, and autistic-likebehavior. There is no effective treatment for fragile X syndrome.

The present invention contemplates the treatment of mild mentalretardation, moderate mental retardation, severe mental retardation,profound mental retardation, and mental retardation severityunspecified. Such mental retardation may be, but is not required to be,associated with chromosomal changes, (for example Down Syndrome due totrisomy 21), heredity, pregnancy and perinatal problems, and othersevere mental disorders. This invention provides methods andcompositions for treating mental retardation using a α5-containingGABA_(A) receptor positive allosteric modulator, such as one selectedfrom the compounds or pharmaceutically acceptable salts, hydrates,solvates, polymorphs, isomers, or combinations thereof as describedherein. In certain embodiments, treatment comprises preventing orslowing the progression of mental retardation. In certain embodiments,treatment comprises alleviation, amelioration, or slowing theprogression of one or more symptoms associated with mental retardation.In certain embodiments, the symptom to be treated is cognitivedeficit/impairment. For example, methods and compositions of thedisclosure can be used to improve the motor/cognitive impairmentssymptomatic of mental retardation. In some embodiments of the invention,there is provided a method of preserving or improving cognitive functionin a subject with mental retardation, the method comprising the step ofadministering to said subject a therapeutically effective amount of acompound of the invention or a pharmaceutically acceptable salt,hydrate, solvate, polymorph, isomer, or combination thereof.

Several animal models have been developed for mental retardation. Forexample, a knockout mouse model has been developed for Fragile Xsyndrome. Fragile X syndrome is a common form of mental retardationcaused by the absence of the FMR1 protein, FMRP. Two homologs of FMRPhave been identified, FXR1P and FXR2P. FXR2P shows high expression inbrain and testis, like FMRP. Both Fxr2 and Fmr1 knockout mice, andFmr1/Fxr2 double knockout mice are believed to be useful models formental retardation such as Fragile X syndrome. See, Bontekoe C. J. M. etal. Hum. Mol. Genet. 2002, 11 (5): 487-498. The efficacy of the methodsand compositions of this invention in treating mental retardation, orcognitive deficit/impairment associated with mental retardation, may beassessed in the these mouse models and other animal models developed formental retardation, as well as human subjects with mental retardation,using a variety of cognitive tests known in the art, as discussedherein.

Compulsive Behavior (Obsessive-Compulsive Disorder)

Obsessive compulsive disorder (“OCD”) is a mental condition that is mostcommonly characterized by intrusive, repetitive unwanted thoughts(obsessions) resulting in compulsive behaviors and mental acts that anindividual feels driven to perform (compulsion). Current epidemiologicaldata indicates that OCD is the fourth most common mental disorder in theUnited States. Some studies suggest the prevalence of OCD is between oneand three percent, although the prevalence of clinically recognized OCDis much lower, suggesting that many individuals with the disorder maynot be diagnosed. Patients with OCD are often diagnosed by apsychologist, psychiatrist, or psychoanalyst according to the Diagnosticand Statistical Manual of Mental Disorders, 4th edition text revision(DSM-IV-TR) (2000) diagnostic criteria that include characteristics ofobsessions and compulsions. Characteristics of obsession include: (1)recurrent and persistent thoughts, impulses, or images that areexperienced as intrusive and that cause marked anxiety or distress; (2)the thoughts, impulses, or images are not simply excessive worries aboutreal-life problems; and (3) the person attempts to ignore or suppresssuch thoughts, impulses, or images, or to neutralize them with someother thought or action. The person recognizes that the obsessionalthoughts, impulses, or images are a product of his or her own mind, andare not based in reality. Characteristics of compulsion include: (1)repetitive behaviors or mental acts that the person feels driven toperform in response to an obsession, or according to rules that must beapplied rigidly; (2) the behaviors or mental acts are aimed atpreventing or reducing distress or preventing some dreaded event orsituation; however, these behaviors or mental acts are not actuallyconnected to the issue, or they are excessive.

Individuals with OCD typically perform tasks (or compulsion) to seekrelief from obsession-related anxiety. Repetitive behaviors such ashandwashing, counting, checking, or cleaning are often performed withthe hope of preventing obsessive thoughts or making them go away.Performing these “rituals,” however, only provides temporary relief.People with OCD may also be diagnosed with a spectrum of other mentaldisorders, such as generalized anxiety disorder, anorexia nervosa, panicattack, or schizophrenia.

The dysfunction in neuronal communication is considered one of theunderlying causes for obsession disorder (Myrrhe van Spronsen and CasperC. Hoogenraad, Curr. Neurol. Neurosci. Rep. 2010, 10, 207-214). Studiessuggest that OCD may be related to abnormal levels of a neurotransmittercalled serotonin. The first-line treatment of OCD consists of behavioraltherapy, cognitive therapy, and medications. Medications for treatmentinclude serotonin reuptake inhibitors (SRIs) such as paroxetine(Seroxat™ Paxil®, Xetanor™, ParoMerck™, Rexetin™), sertraline (Zoloft®,Stimuloton™) fluoxetine (Prozac®, Bioxetin™), escitalopram (Lexapro®),and fluvoxamine (Luvox®) as well as the tricyclic antidepressants, inparticular clomipramine (Anafranil®). Benzodiazepines are also used intreatment. As much as 40 to 60% of the patients, however, fail toadequately respond to the SRI therapy and an even greater proportion ofpatients fail to experience complete remission of their symptoms.

The invention provides methods and compositions for treating OCD using aα5-containing GABA_(A) receptor agonist (e.g., a α5-containing GABA_(A)receptor positive allosteric modulator), such as one selected from thecompounds or pharmaceutically acceptable salts, hydrates, solvates,polymorphs, isomers, or combinations thereof as described herein. Incertain embodiments, treatment comprises preventing or slowing theprogression of OCD. In certain embodiments, treatment comprisesalleviation, amelioration, or slowing the progression of one or moresymptoms associated with OCD. In certain embodiments, the symptom to betreated is cognitive impairment or cognitive deficit. For example,methods and compositions of the disclosure can be used to treat thecognitive deficits in OCD, and/or to improve cognitive function inpatients with OCD. In some embodiments of the invention, there isprovided a method of preserving or improving cognitive function in asubject with OCD, the method comprising the step of administering tosaid subject a therapeutically effective amount of a compound of theinvention or a pharmaceutically acceptable salt, hydrate, solvate,polymorph, isomer, or combination thereof.

A quinpirole-sensitized rat model has been developed for OCD. Thecompulsive checking behavior of the quinpirole-sensitized rats issubject to interruption, which is an attribute characteristic of OCDcompulsions. In addition, a schedule-induced polydipsia (SIP) rodentmodel of obsessive-compulsive disorder was used to evaluate the effectsof the novel 5-HT2C receptor agonist WAY-163909. See, e.g.,Rosenzweig-Lipson S. et al. Psychopharmacology (Berl) 2007, 192, 159-70.The efficacy of the methods and compositions of this invention intreating OCD, or cognitive impairment or cognitive deficits associatedwith OCD, may be assessed in the above animal models and other animalmodels developed for OCD, as well as human subjects with OCD, using avariety of cognitive tests known in the art, as discussed herein.

Substance Addiction

Substance addiction (e.g., drug substance addiction, alcohol substanceaddiction) is a mental disorder. The substance addiction is nottriggered instantaneously upon exposure to substance of abuse. Rather,it involves multiple, complex neural adaptations that develop withdifferent time courses ranging from hours to days to months (Kauer J. A.Nat. Rev. Neurosci. 2007, 8, 844-858). The path to substance addictiongenerally begins with the voluntary use of one or more controlledsubstances, such as narcotics, barbiturates, methamphetamines, alcohol,nicotine, and any of a variety of other such controlled substances. Overtime, with extended use of the controlled substance(s), the voluntaryability to abstain from the controlled substance(s) is compromised dueto the effects of prolonged use on brain function, and thus on behavior.As such, substance addiction generally is characterized by compulsivesubstance craving, seeking and use that persist even in the face ofnegative consequences. The cravings may represent changes in theunderlying neurobiology of the patient which likely must be addressed ina meaningful way if recovery is to be obtained. Substance addiction isalso characterized in many cases by withdrawal symptoms, which for somesubstances are life threatening (e.g., alcohol, barbiturates) and inothers can result in substantial morbidity (which may include nausea,vomiting, fever, dizziness, and profuse sweating), distress, anddecreased ability to obtain recovery. For example, alcoholism, alsoknown as alcohol dependence, is one such substance addiction. Alcoholismis primarily characterized by four symptoms, which include cravings,loss of control, physical dependence and tolerance. These symptoms alsomay characterize substance addictions to other controlled substances.The craving for alcohol, as well as other controlled substances, oftenis as strong as the need for food or water. Thus, an alcoholic maycontinue to drink despite serious family, health and/or legalramifications.

Recent work exploring the effects of abusing alcohol, centralstimulants, and opiates on the central nervous system (CNS) havedemonstrated a variety of adverse effects related to mental health,including substance-induced impairments in cognition. See, Nyberg F.Cognitive Impairments in Drug Addicts, Chapter 9. In severallaboratories and clinics substantial damages of brain function are seento result from these drugs. Among the harmful effects of the abusingdrugs on brain are those contributing to accelerated obsolescence. Anobservation that has received special attention during recent years isthat chronic drug users display pronounced impairment in brain areasassociated with executive and memory function. A remarkedneuroadaptation caused by addictive drugs, such as alcohol, centralstimulants and opiates involves diminished neurogenesis in thesubgranular zone (SGZ) of the hippocampus. Indeed, it has been proposedthat decreased adult neurogenesis in the SGZ could modify thehippocampal function in such a way that it contributes to relapse and amaintained addictive behavior. It also raises the possibility thatdecreased neurogenesis may contribute to cognitive deficits elicited bythese abusing drugs.

The invention provides methods and compositions for treating substanceaddiction using a α5-containing GABA_(A) receptor positive allostericmodulator, such as one selected from the compounds or pharmaceuticallyacceptable salts, hydrates, solvates, polymorphs, isomers, orcombinations thereof as described herein. In certain embodiments,treatment comprises preventing or slowing the progression of substanceaddiction. In certain embodiments, treatment comprises alleviation,amelioration, or slowing the progression of one or more symptomsassociated with substance addiction. In certain embodiments, the symptomto be treated is cognitive impairment. For example, methods andcompositions of the disclosure can be used to treat the cognitiveimpairment and/or to improve cognitive function in patients withsubstance addiction. In some embodiments of the invention, there isprovided a method of preserving or improving cognitive function in asubject with substance addiction, the method comprising the step ofadministering to said subject a therapeutically effective amount of acompound of the invention or a pharmaceutically acceptable salt,hydrate, solvate, polymorph, isomer, or combination thereof.

Several animal models have been developed to study substance addiction.For example, a genetically selected Marchigian Sardinianalcohol-preferring (msP) rat models was developed to study theneurobiology of alcoholism. See, Ciccocioppo R. et al. Substanceaddiction Biology 2006, 11, 339-355. The efficacy of the methods andcompositions of this invention in treating substance addiction, orcognitive impairment associated with substance addiction, may also beassessed in animal models of substance addiction, as well as humansubjects with substance addiction, using a variety of cognitive testsknown in the art, as discussed herein.

Research Domain Criteria (RDoC)

The invention further provides methods and compositions for treatingimpairment in neurological disorders and neuropsychiatric conditionsusing a α5-containing GABA_(A) R positive allosteric modulator or apharmaceutically acceptable salt, hydrate, solvate, polymorph, isomer,or combination thereof as described herein. In certain embodiments,treatment comprises alleviation, amelioration or slowing theprogression, of one or more symptoms associated with such impairment. Inanother aspect of the invention, there is provided methods andcompositions for preserving or improving cognitive function in a subjectin need thereof using a compound of the invention or a pharmaceuticallyacceptable salt, hydrate, solvate, polymorph, isomer, or combinationthereof.

Research Domain Criteria (RDoC) are expected to augment clinicalcriteria, such as DSM and ICD, for diagnosis of disease and disordersaffecting the nervous system (see, e.g., Am. J. Psychiatry 167:7(2010)). The RDoC is intended to provide classification based ondiscoveries in genomics and neuroscience as well as clinicalobservation. The high expression of α5-containing GABA_(A) receptors inspecific neural circuits in the nervous system could be therapeutictargets for neural circuit dysfunction identified under RDoC.

Assays for GABA_(A) α5 Subunit Binding and Receptor Positive AllostericModulator Activity

The affinity of test compounds for a GABA_(A) receptor comprising theGABA_(A) α5 subunit may be determined using receptor binding assays thatare known in the art. See, e.g., U.S. Pat. Nos. 7,642,267 and 6,743,789,which are incorporated herein by reference.

The activity of the test compounds as a α5-containing GABA_(A) Rpositive allosteric modulator may be tested by electrophysiologicalmethods known in the art. See, e.g., U.S. Pat. No. 7,642,267 andGuidotti et al., Psychopharmacology 180: 191-205, 2005. Positiveallosteric modulator activity may be tested, for examples, by assayingGABA-induced chloride ion conductance of GABA_(A) receptors comprisingthe GABA_(A) α5 subunit. Cells expressing such receptors may be exposedto an effective amount of a compound of the invention. Such cells may becontacted in vivo with compounds of the invention through contact with abody fluid containing the compound, for example through contact withcerebrospinal fluid. In vitro tests may be done by contacting cells witha compound of the invention in the presence of GABA. IncreasedGABA-induced chloride conductance in cells expressing GABA_(A) receptorscomprising the GABA_(A) α5 subunit in the presence of the test compoundwould indicate positive allosteric modulator activity of said compound.Such changes in conductance may be detected by, e.g., using avoltage-clamp assay performed on Xenopus oocytes injected with GABA_(A)receptor subunit mRNA (including GABA_(A) α5 subunit RNA), HEK 293 cellstransfected with plasmids encoding GABA_(A) receptor subunits, or invivo, ex vivo, or cultured neurons.

It will be understood by one of ordinary skill in the art that themethods described herein may be adapted and modified as is appropriatefor the application being addressed and that the methods describedherein may be employed in other suitable applications, and that suchother additions and modifications will not depart from the scope hereof.

This invention will be better understood from the Examples which follow.However, one skilled in the art will readily appreciate that thespecific methods and results discussed are merely illustrative of theinvention as described more fully in the embodiments which followthereafter.

Example 1: Synthesis of Compound 1

To a stirred mixture of 5-methoxy-2-nitroaniline (5 g, 29.7 mmol) in HCl(conc. 39 mL) at 0° C. was added drop wise a solution of NaNO₂ (2.05 g,29.7 mmol) in H₂O (19 mL). The internal temperature was kept below 10°C. After addition, the mixture was stirred at room temperature for 1 h.The diazonium salt was collected by filtration, and was used in the nextstep. To the diazonium salt in a crystallization dish under faststirring at room temperature was added drop wise a solution of NaN₃(1.93 g, 29.6 mmol) in H₂O (7 mL). After gas evolution stopped (3 h), itwas filtered. The collected solid was re-crystallized from MeOH to give4.342 g (yield 75% for 2 steps) of the product 13 as a yellow solid. Toa mixture of the phenylazide 13 (1.94 g, 10 mmol) and diethyl1,3-acetone-diacrboxylate (2.20 mL, 12 mmol) in EtOH (40 mL) at roomtemperature was added Et₃N (1.67 mL, 12 mmol). After the mixture wasstirred at room temperature for 60 h, the initial suspension turned intoa clear yellow solution. The solution was concentrated under vacuum andthe residue was purified by chromatography (RediSep 24 g silica-gelcolumn, 10% to 40% EtOAc in hexanes) to give 2.905 g of triazole 14 as ayellow solid. MS: [M+1]=379.

The above triazole 14 (2.95 g, 7.66 mmol) in EtOH (50 mL) with Pd/C (10wt %, 407 mg, 0.38 mmol) was stirred under H₂ (balloon) for 24 h. It wasfiltered through Celite. The filtrate was concentrated and the residuewas purified by chromatography (RediSep 24 g silica-gel column, 10% to50% EtOAc in hexanes) to give 2.453 g of aniline 15 as a white solid.(70% yield for two steps.) MS: [M+1]=349.

Compound 15 (2.45 g, 7.03 mmol) and catalytic amount of p-TsOH.H₂O (24mg) in p-xylene (30 mL) were heated in a 140° C. oil bath overnight. Themixture was cooled and filtered. The solid was washed with cold EtOAc.After drying, it gave 1.88 g (88% yield) of the lactam 16. MS:[M+1]=303.

To a suspension of the lactam ester 16 (837 mg, 2.77 mmol) in THE (20mL) at room temperature was add LiBH₄ (2 M in THF, 1.39 mL, 2.78 mmol).After the mixture was stirred at room temperature for 60 h, more LiBH₄(2 M in THF, 0.28 mL, 0.56 mmol) was added and it was stirred at roomtemperature for 24 additional h. A mixture of EtOAc/EtOH (10 mL/10 mL)was added to the reaction and it was concentrated in vacuo. The residuewas taken up in EtOAc/CH₂Cl₂/MeOH and loose silica gel was added. Aftervolatile solvents were evaporated, the solid was loaded onto a RediSep24 g silica-gel column. Chromatography (solvent A: EtOAc, solvent B:10:1 v/v CH₂Cl₂/MeOH; gradient eluent: A to B) gave 540 mg (75% yield)of the alcohol 17 as white solid. MS: [M+1]=261.

To a solution of the alcohol 17 (105.4 mg, 0.40 mmol) and CBr₄ (336 mg,1.01 mmol) in DMF (3 mL) was slowly added a solution of PPh₃ (255 mg,0.97 mmol) in DMF (1 mL) over 20 min. After addition, TLC showed thereaction went completion. Water was added to quench the reaction and themixture was extracted with EtOAc thrice. The combined extracts werewashed sequentially with H₂O, brine and dried over Na₂SO₄. Filtrationand concentration gave the crude product. Chromatography (RediSep 12 gsilica-gel column, CH₂Cl₂ to 30% EtOAc in CH₂Cl₂) gave 439.2 mg of amixture of the bromide 18 ([M+1]=324) and Ph₃PO. The above mixture (439mg) in EtOAc/EtOH (8 mL/8 mL) with Pd/C (10 wt %, 200 mg, 0.19 mmol) wasstirred under H₂ (balloon) for 2 h, then was filtered through Celite.The filtrate was concentrated and residue was purified by chromatography(RediSep 12 g silica-gel column, solvent A: 1:1 v/v CH₂Cl₂/hexanes,solvent B: EtOAc; gradient eluent: A to B) to give 99 mg (˜80% yield for2 steps) of product 19 as a white solid. MS: [M+1]=245.

In a separate flask, 1,2,3-triazole (55.3 mg, 0.80 mmol) in CH₃CN (1 mL)at 0° C. was treated with i-Pr₂NEt (146 μL, 0.84 mmol), followed byPOCl₃ (23 μL, 0.25 mmol). The solution was stirred at 0° C. for 2 h. Thelactam 19 was added in one lot and the resulting suspension was heatedin an 80° C. oil bath for 20 h. Water was added to quench the reaction.It was extracted with EtOAc thrice. The combined extracts were washedwith brine and dried over Na₂SO₄. Filtration and concentration gave 48.8mg of the crude product 20, which was used directly in the next step. Asolution of KO-t-Bu (37.2 mg, 0.33 mmol) in DMF (0.5 mL) was cooled to−50° C. Ethyl isocyanoacetate (40 μL, 0.36 mmol) was added drop wise.The mixture was stirred at −50° C. for 1 h. The above crude product 20in DMF (1 mL) was added drop wise. The mixture was allowed to warm to10° C. and stirred at 10° C. for 1 h. Saturated NH₄Cl aqueous solutionwas added and it was extracted with EtOAc thrice. The combined extractswere washed sequentially with water, brine and dried over Na₂SO₄.Filtration and concentration gave the crude product.

Chromatography (RediSep 12 g silica-gel column, solvent A: 1:1 v/vCH₂Cl₂/hexanes, solvent B: EtOAc; gradient eluent: 20% to 80% B in A) togive 15 mg (21% yield for 2 steps) of Compound 1 (Example 1) as anoff-white solid. MS: [M+1]=340. ¹H-NMR (500 MHz, CDCl3) δ: 7.74 (s, 1H),7.63 (d, 1H, J=3 Hz), 7.51 (d, 1H, J=8.5 Hz), 7.14 (dd, 1H, J=3.0, 8.5Hz), 4.44 (q, 2H, J=7.0 Hz), 3.95 (s, 3H), 2.44 (s, 3H), 1.45 (t, 3H,J=7.0 Hz).

Example 2: Synthesis of Compound 2

Compound of Example 2 was synthesized in an analogous synthetic route asthat described for Example 1, using 5-fluoro-2-nitro-aniline as thestarting material to give Compound 2 as a light brown solid: MS:[M+1]=328. ¹H-NMR (500 MHz, CDCl3) δ: 7.90 (br dd, 1H, J=2.5, 8.5 Hz),7.77 (s, 1H), 7.62 (br dd, 1H, J=5.0, 9.0 Hz), 7.35 (m, 1H), 4.45 (q,2H, J=7.0 Hz), 2.45 (s, 3H), 1.45 (t, 3H, J=7.0 Hz).

Example 3: Synthesis of Compound 3

Compound of Example 3 was synthesized in an analogous synthetic route asthat described for Example 1, using 2-nitro-aniline as the startingmaterial to give Compound 3 as a light yellow solid: MS: [M+1]=310;¹H-NMR (500 MHz, CDCl3) δ: 8.161 (br d, 1H, J=8.5 Hz), 7.81 (s, 1H),7.66 (m, 3H), 4.45 (q, 2H, J=7.0 Hz), 2.45 (s, 3H), 1.46 (t, 3H, J=7.0Hz).

Example 4: Synthesis of Compound 110

Acetamide oxime was azeotroped three times in toluene before use. To asuspension of acetamide oxime (30 mg, 0.4 mmol) in THF (1 mL) was addedNaH 60% in oil dispersion (16 mg, 0.4 mmol). The suspension was stirredat room temperature for 15 min. The ester compound 2 (65 mg, 0.2 mmol)was added. The vial containing the ester was rinsed with THE (1 mL)which was added to the reaction mixture. The resulting brown suspensionwas stirred at room temperature for 30 mins. then heated at 70° C. for 2h 30 min. The suspension was quenched with MeOH. The solvent wasevaporated and the crude oil was purified by chromatography (RediSep 4 gsilica-gel column, eluted with 70% EtOAc in Hexanes) to give 28 mg (41%yield) of product. MS: [M+1]=338. H¹NMR (CDCl₃) δ 7.92 (1H, dd, J=2.5,8.5 Hz), 7.90 (1H, s), 7.67 (1H, dd, J=4.5, 9.5 Hz), 7.38 (1H, m), 2.51(3H, s), 2.46 (3H, s).

Example 5: Synthesis of Compound 167

The compound was prepared analogously from Compound 1 to give Compound167: MS: [M+1]=350. H¹NMR (CDCl₃) δ 7.87 (1H, s), 7.65 (1H, d, J=3 Hz),7.55 (1H, d, J=9 Hz), 7.17 (1H, dd, J=2.5, 9 Hz), 3.96 (3H, s), 2.5 (3H,s), 2.45 (3H, s).

Example 6: Synthesis of Compound 4

To a solution of compound 17 prepared as in Example 1 (260 mg) in DMSO(4 mL) and CH₂Cl₂ (6 mL) was added Et₃N (0.7 mL, 5 mmol), followed byPy.SO₃ (398 mg, 2.5 mmol). It was stirred at room temperature for 1 h.The reaction mixture was poured into water and extracted with EtOActhrice. The combined extracts were washed sequentially with H₂O, brineand dried over Na₂SO₄. Filtration and concentration gave 198.5 mg of thecrude aldehyde 21, which was used without further purification. To asuspension of aldehyde 21 (198.5 mg, 0.77 mmol) in THE (10 mL) at 0° C.was added drop wise PhMgBr (1 M in THF, 1.54 mL, 1.54 mmol). It wasstirred at 0° C. for 30 min. Saturated NH₄Cl aqueous solution was addedand it was extracted with EtOAc thrice.

The combined extracts were washed with brine and dried over Na₂SO₄.Filtration and concentration gave 252.9 mg of the benzyl alcohol 22 as abrown foamy solid. This was used in the next step without furtherpurification. To a solution of the above crude alcohol 22 in CH₂Cl₂ (8mL) with Et₃SiH (0.60 mL, 3.76 mmol) was added TFA (0.64 mL, 8.27 mmol).The reaction solution was stirred at room temperature for 4 h. Afterconcentration, the residue was purified by chromatography (RediSep 12 gsilica-gel column, 20% to 80% EtOAc in hexanes) to give 34.1 mg (yield12% for four steps) of the reduced product 23 as white foamy solid. MS:[M+1]=321.

In a separate flask, a solution of 1,2,4-triazole (27 mg, 0.39 mmol) inCH₃CN (0.5 mL) at 0° C. was treated with i-Pr₂NEt (72 μL, 0.41 mmol),followed by POCl₃ (11 μL, 0.12 mmol). The mixture was stirred at 0° C.for 2 h. The lactam material 23 (32.2 mg, 0.1 mmol, solid) was added inone lot to the reaction mixture and it was heated in an 80° C. oil bathfor 20 h. The mixture was cooled to room temperature and creamy solidprecipitate was observed. Water (0.5 mL) was added and it was stirred atroom temperature for 5 min. The solid precipitate was collected byfiltration, and washed with 0.5 mL of water, followed by drying underhigh vacuum to give 15.8 mg (yield 42%) of the adduct 24 as a off-whitefluffy solid. MS: [M+1]=372. A solution of KO-t-Bu (9.5 mg, 85 μmol) inDMF (0.5 mL) was cooled to −50° C. Ethyl isocyanoacetate (10.4 μL, 95μmol) was added drop wise. The resulting mixture was stirred at −50° C.for 1 h. The triazole amidine 24 (15.8 mg, 42 μmol, solid) was added inone lot. The stirred mixture was allowed to warm up to 10° C. in 1 h andkept at 10° C. for 1 h. Saturated NH₄Cl aqueous solution was added andit was extracted with EtOAc thrice. The combined extracts were washedsequentially with H₂O, brine and dried over Na₂SO₄. Filtration andconcentration gave the crude product. Chromatography (RediSep 4 gsilica-gel column. Solvent A: 1:1 v/v CH₂Cl₂/hexanes, solvent B: EtOAc;gradient eluent: A to 50% B in A) gave 16.8 mg (yield 95%) of thecompound of Example 6 as a white solid. MS: [M+1]=416. ¹H-NMR (500 MHz,CDCl3) δ: 7.74 (s, 1H), 7.63 (d, 1H, J=3.0 Hz), 7.50 (d, 1H, J=9.0 Hz),7.30 (br d, 2H, J=7.0 Hz), 7.29 (br d, 2H, 7.5 Hz), 7.20 (m, 1H), 7.13(dd, 1H, J=2.5, 9.0 Hz), 4.41 (q, 2H, J=7.5 Hz), 4.17 (s, 2H), 3.95 (s,3H), 1.43 (t, 3H, 7.5 Hz).

Example 7: Synthesis of Compound 5

Compound of Example 7 was synthesized in an analogous synthetic route asthat described for Example 6, using 2-nitro-aniline as the startingmaterial to give Compound 5 as a brown solid: MS: [M+1]=386. ¹H-NMR (500MHz, CDCl3) δ: 8.16 (br d, 1H, J=7.0 Hz), 7.81 (s, 1H), 7.60-7.68 (m,3H), 7.34 (br d, 2H, J=8.0 Hz), 7.29 (br d, 2H, J=7.0 Hz), 7.20 (m, 1H),4.42 (q, 2H, J=7.0 Hz), 4.18 (s, 2H), 1.44 (t, 3H, J=7.0 Hz).

Example 8: Synthesis of Compound 6

Compound of Example 8 was synthesized in an analogous synthetic route asthat described for Example 6, using 5-fluoro-2-nitro-aniline as thestarting material to give compound 8 as a brown solid: MS: [M+1]=404.¹H-NMR (500 MHz, CDCl3) δ: 7.90 (dd, 1H, J=3.5, 8.5 Hz), 7.77 (s, 1H),7.61 (dd, 1H, J=5.0, 10.5 Hz), 7.28-7.37 (m, 5H), 7.21 (m, 1H), 4.43 (q,2H, J=7.0 Hz), 4.17 (s, 2H), 1.44 (t, 3H, J=7.0 Hz).

Example 9: Synthesis of Compound 44

Compound of Example 9 was synthesized in an analogous synthetic route asthat described for Example 6, using 5-fluoro-2-nitro-aniline as thestarting material to give the compound of Example 9 as a brownish solid:MS: [M+1]=418. ¹H-NMR (500 MHz, CDCl3) δ: 7.89 (br d, 1H, J=9.5 Hz),7.76 (s, 1H), 7.60 (dd, 1H, J=5.5, 10.0 Hz), 7.35 (br t, 1H, J=6.0 Hz),7.22 (br d, 2H, J=8.5 Hz), 7.09 (br d, 2H, J=7.5 Hz), 4.43 (q, 2H, J=7.5Hz), 4.12 (s, 2H), 2.30 (s, 3H), 1.44 (t, 3H, J=7.5 Hz).

Example 10: Synthesis of Compound 45

Compound of Example 10 was synthesized in an analogous synthetic routeas that described for Example 6, using 5-fluoro-2-nitro-aniline as thestarting material to give the compound of Example 10 as a brownishsolid: MS: [M+1]=438. ¹H-NMR (500 MHz, CDCl3) δ: 7.90 (dd, 1H, J=3.0,8.0 Hz), 7.77 (s, 1H), 7.61 (dd, 1H, J=5.0, 9.0 Hz), 7.36 (m, 1H), 7.25(br s, 4H), 4.42 (q, 2H, J=7.0 Hz), 4.14 (s, 2H), 1.44 (t, 3H, J=7.0Hz).

Example 11: Synthesis of Compound 46

Compound of Example 11 was synthesized in an analogous synthetic routeas that described for Example 6, using 5-fluoro-2-nitro-aniline as thestarting material to give the compound of Example 11 as a yellowishsolid: MS: [M+1]=422. ¹H-NMR (500 MHz, CDCl3) δ: 7.90 (dd, 1H, J=3.0,8.5 Hz), 7.77 (s, 1H), 7.61 (dd, 1H, J=5.0, 9.0 Hz), 7.36 (m, 1H), 7.28(m, 2H), 6.96 (m, 2H), 4.42 (q, 2H, J=7.5 Hz), 4.14 (s, 2H), 1.44 (t,3H, J=7.0 Hz).

Example 12: Synthesis of Compound 47

Compound of Example 12 was synthesized in an analogous synthetic routeas that described for Example 6, using 2-nitro-aniline as the startingmaterial to give the compound of Example 12 as a yellowish solid: MS:[M+1]=420. ¹H-NMR (500 MHz, CDCl3) δ: 8.16 (br d, 1H, J=7.0 Hz), 7.80(s, 1H), 7.64 (m, 3H), 7.25 (m, 4H), 4.41 (q, 2H, J=7.0 Hz), 4.14 (s,2H), 1.44 (t, 3H, J=8.0 Hz).

Example 13: Synthesis of Compound 109

Acetamide oxime (50 mg, 0.67 mmol) was azeotroped with toluene 3 times.THF (5 mL) was added, then NaH 60% in oil dispersion (25 mg, 0.62 mmol).The suspension was stirred at room temperature for 30 min. 2 mL of thissuspension was added to ester compound 6 (40 mg, 0.099 mmol) and theresulting solution was heated at 70° C. for 3 h. The solution wasquenched with water. The solution was extracted with EtOAc (3×). Thecombined organic phases were washed with brine, dried over MgSO₄.Filtration and concentration gave the crude product. Chromatography(RediSep 12 g silica-gel column. Eluted with 50% EtOAc in Hexanes) gave6 mg (yield 20%) of the product Compound 109 as yellow solid. MS:[M+1]=414). H¹NMR (CDCl₃) δ 7.93 (1H, dd, J=3, 8.5 Hz), 7.89 (1H, s),7.65 (1H, dd, J=5.5, 9 Hz), 7.38 (1H, m), 7.23 (5H, m), 4.2 (2H, s),2.50 (3H, s).

Example 14: Synthesis of Compound 7

To a stirred mixture of 5-methoxy-2-nitroaniline (5 g, 29.7 mmol) in HCl(conc. 12.9 mL) at 0° C. was added drop wise a solution of NaNO₂ (2.05g, 29.7 mmol) in H₂O (8 mL). The internal temperature was kept below 5°C. After addition, the mixture was allowed to warm up to roomtemperature in 1 h. The mixture was cooled to 0° C. and a solution ofSnCl₂.2H₂O (20.13 g, 89.2 mmol) in HCl (conc. 13 mL) was added slowlydropwise. After addition, it was stirred at room temperature for 2 h.The resulting yellow solid was collected by filtration and washed withcold (0° C.) 6 N HCl. After drying in vacuum oven, it gave 3.245 g(yield 50%) of brown solid as aryl hydrazine 25. MS: [M+H₂O+Na]=224. Ina separate flask, a mixture of diethyl 1,3-acetonediacrboxylate (2.426g, 12 mmol) and diethoxymethyl acetate (1.946 g, 12 mmol) was heatedunder microwave radiation at 100° C. for 1 h. The reaction mixture wasconcentrated in vacuo, and residual volatile component was co-distilledoff with toluene (5 ml) in vacuo to give condensation product 26, whichwas used directly in the next step.

Product 26 from above was dissolved in EtOH (30 mL). Molecular sieves (4Å, 2 g) and hydrazine hydrochloride 25 (2.19 g, 10 mmol) were added. Thesuspension was stirred at room temperature for 24 h. It was filteredthrough Celite and the solid was washed with EtOAc (10 mL×3). Thefiltrate was concentrated. The residue was purified by chromatography(RediSep 40 g silica-gel column, 10% to 40% EtOAc in hexanes) to give2.091 g of pyrrole 27 which was used without further purification in thenext step. MS: [M+1]=378.

The above nitro group on 27 (2.09 g, 5.5 mmol) was reduced in EtOH (40mL) with Pd/C (10 wt %, 295 mg, 0.28 mmol) under H₂ (balloon) for 18 h.The mixture was filtered through Celite. The filtrate was concentratedand the residue was purified by chromatography (RediSep 24 g silica-gelcolumn, hexanes to 50% EtOAc in hexanes) to give 1.127 g of theun-cyclized product 28 as a yellow sticky oil ([M+1]=348), plus 154 mgof cyclized product 29 as a gray solid (MS: [M+1]=302). The un-cyclizedaniline 28 (1.127 g, 3.2 mmol) in p-xylene (20 mL) was treated withcatalytic amount of p-TsOH. H₂O (15 mg) in a 140° C. oil bath for 20 h.The reaction mixture was cooled, concentrated, and the residue wastriturated with cold (0° C.) EtOAc. Filtration gave 559 mg of the lactamproduct 29 as a yellow solid. The total weight of the lactam product 29combined is 713 mg (24% for 3 steps). MS: [M+1]=302.

To a suspension of the ester 29 (566 mg, 1.88 mmol) in CH₂Cl₂ (35 mL) at−78° C. was added Dibal-H (1 M in hexane, 6.60 mL, 6.60 mmol). Thesuspension was stirred for 10 min at −78° C. The cold bath was removedand it was stirred for 20 min while the temperature rose to roomtemperature. At this point, TLC showed ˜80% reaction completion. It wascooled to −78° C. and more Dibal-H (1 M in hexane, 1.0 mL, 1.0 mmol) wasadded. After stirring at −78° C. for 30 min, LCMS showed the reactionproceeded to completion. The reaction was quenched by addition ofRochelle's salt aqueous solution (20%) followed by EtOAc. It wasvigorously stirred at room temperature until it became a clear two-layermixture. The layers were separated and the aqueous layer was extractedwith EtOAc thrice. The combined organic phase was washed with brine anddried over Na₂SO₄. Filtration and concentration gave 480 mg of the crudealcohol 30 as a slightly yellow solid. MS: [M+1]=260.

To a solution of alcohol 30 (200 mg, 0.77 mmol) and CBr₄ (640 mg, 1.93mmol) in DMF (8 mL) was added a solution of PPh₃ (486 mg, 1.85 mmol) inDMF (2 mL) slowly in 30 min. After addition, it was stirred at roomtemperature for 30 min. Water was added to quench the reaction and themixture was extracted with EtOAc thrice. The combined extracts werewashed sequentially with H₂O, brine and dried over Na₂SO₄. Filtrationand concentration gave the crude product. Chromatography (RediSep 12 gsilica-gel column, solvent A: 1:1 v/v CH₂Cl₂/hexanes, solvent B: EtOAc;gradient eluent: 10% to 40% B in A) gave 221 mg of a mixture of thebromide 31 and Ph₃PO.

The above mixture in EtOAc/EtOH (8 mL/8 mL) with Pd/C (10 wt %, 200 mg,0.19 mmol) was stirred under H₂ (balloon) for 1 h. It was filteredthrough Celite. The filtrate was concentrated and residue was purifiedby chromatography (RediSep 12 g silica-gel column, solvent A: 1:1 v/vCH₂Cl₂/hexanes, solvent B: EtOAc; gradient eluent: 10% to 40% B in A) togive 146 mg of a mixture of the reduction product 32 ([M+1]=244) andPh₃PO.

In a separate flask, 1,2,4-triazole (81 mg, 1.17 mmol) in CH₃CN (1 mL)at 0° C. was treated with i-Pr₂NEt (214 μL, 1.23 mmol), followed byPOCl₃ (34 μL, 0.36 mmol). The solution was stirred at 0° C. for 2 h. Thelactam 32 (˜60% purity by LCMS) was added in one lot and the resultingsuspension was heated in an 80° C. oil bath for 18 h. Water was added toquench the reaction. It was extracted with EtOAc thrice. The combinedextracts were washed sequentially with H₂O, brine and dried over Na₂SO₄.Filtration and concentration gave 126.6 mg of the crude product 33 as ayellow glue, which was used directly in the next reaction. MS:[M+1]=295. A solution of KO-t-Bu (97 mg, 0.86 mmol) in DMF (1 mL) wascooled to −50° C. Ethyl isocyanoacetate (104 μL, 0.95 mmol) was addeddrop wise. The mixture was stirred at −50° C. for 1 h. The above crudeproduct 33 in DMF (1.5 mL) was added drop wise. The mixture was allowedto warm to 10° C. and stirred at 10° C. for 1 h. Saturated NH₄Cl aqueoussolution was added and it was extracted with EtOAc thrice. The combinedextracts were washed sequentially with water, brine and dried overNa₂SO₄. Filtration and concentration gave the crude product.Chromatography (RediSep 12 g silica-gel column, solvent A: 1:1 v/vCH₂Cl₂/hexanes, solvent B: EtOAc; gradient eluent: 10% to 40% B in A) togive 22 mg of a white solid, which was further purified by preparativeTLC (developed with 1:1 A/B) to give 12.8 mg of the final productCompound 7 (Example 14) as a white solid. MS: [M+1]=339. ¹H-NMR (500MHz, CDCl3) δ: 7.70 (s, 1H), 7.56 (s, 1H), 7.50 (d, 1H, J=3.0 Hz), 7.43(d, 1H, J=8.5 Hz), 7.00 (dd, 1H, J=2.5, 9.5 Hz), 5.29 (br s, 1H), 4.44(q, 2H, J=7.0 Hz), 3.92 (s, 3H), 3.55 (br s, 1H), 2.17 (s, 3H), 1.45 (t,3H, J=7.0 Hz).

Example 15: Synthesis of Compound 8

To a solution of the alcohol 30 (261 mg, 1.0 mmol) which was prepared inExample 14 in DMSO (4 mL) and CH₂Cl₂ (6 mL) was added Et₃N (0.7 mL, 5mmol), followed by Py.SO₃ (398 mg, 2.5 mmol). It was stirred at roomtemperature for 1 h. The reaction mixture was poured into water andextracted with EtOAc thrice. The combined extracts were washedsequentially with H₂O, brine and dried over Na₂SO₄. Filtration andconcentration gave 226 mg of the crude aldehyde 34 as a yellow solid. Itwas used in the next step without purification. MS: [M+1]=258.

To a suspension of the crude aldehyde 34 (202 mg, 0.79 mmol) in THE (10mL) at 0° C. was added drop wise PhMgBr (1 M in THF, 1.58 mL, 1.58mmol). It was stirred at 0° C. for 30 min. Saturated NH₄Cl aqueoussolution was added and it was extracted with EtOAc thrice. The combinedextracts were washed with brine and dried over Na₂SO₄. Filtration andconcentration gave 275 mg of the crude product 35 as a yellow foamysolid, which was used in the next step without purification.

To a solution of the above crude alcohol 35 in CH₂Cl₂ (10 mL) withEt₃SiH (0.66 mL, 4.10 mmol) was added TFA (0.70 mL, 9.02 mmol). Thereaction solution was stirred at room temperature for 1 h. Afterconcentration, the residue was purified by chromatography (RediSep 24 gsilica-gel column, 10% to 50% EtOAc in hexanes) to give 187.8 mg (yield59% for three steps) of the product 36 as a gray solid. MS: [M+1]=320.

In a separate flask, a solution of 1,2,4-triazole (127 mg, 1.83 mmol) inCH₃CN (1.6 mL) at 0° C. was treated with i-Pr₂NEt (336 μL, 1.93 mmol),followed by POCl₃ (53 μL, 0.56 mmol). The mixture was stirred at 0° C.for 2 h. Lactam 36 (150 mg, 0.47 mmol, solid) was added in one lot tothe reaction mixture and it was heated in an 80° C. oil bath for 18 h.The mixture was cooled to room temperature and solid precipitate wasobserved. Water (2.1 mL) was added and it was stirred at roomtemperature for 10 min. Filtration, washing the solid with 2 mL ofwater, followed by drying under high vacuum gave 118.8 mg (yield 69%) ofthe triazole amidine 37 as an off-white fluffy solid. MS: [M+1]=371. Asolution of KO-t-Bu (72 mg, 0.64 mmol) in DMF (2 mL) was cooled to −50°C. Ethyl isocyanoacetate (77 μL, 0.71 mol) was added drop wise. Theresulting mixture was stirred at −50° C. for 1 h. The triazole amidine37 (118.8 mg, 42 μmol, solid) was added in lot. The stirred mixture wasallowed to warm up to 10° C. in 1 h and kept at 10° C. for 1 h.Saturated NH₄Cl aqueous solution was added and it was extracted withEtOAc thrice. The combined extracts were washed sequentially with H₂O,brine and dried over Na₂SO₄. Filtration, concentration, thenchromatography (RediSep 12 g silica-gel column. solvent A: 1:1 v/vCH₂Cl₂/hexanes, solvent B: EtOAc; gradient eluent: A to 40% B in A) gave125.1 mg (yield 94%) of Compound 8 as a white solid. MS: [M+1]=415.¹H-NMR (500 MHz; CDCl₃) δ: 7.72 (s, 1H), 7.54 (s, 1H), 7.51 (br s, 1H),7.44 (br d, 1H, J=9.5 Hz), 7.29 (br d, 2H, J=7.5 Hz), 7.20 (m, 3H), 7.01(br d, 1H, J=7.5 Hz), 5.30 (br s, 1H), 4.38 (q, 2H, J=7.0 Hz), 3.92 (brs, 5H), 3.54 (br s, 1H), 1.41 (t, 3H, J=7.0 Hz).

Example 16: Synthesis of Compound 9

LiOH (1.09 g, 45.5 mmol) was added to a stirring solution of ester 16(prepared in Example 1) (2.75 g, 9.10 mmol) in THE (24 mL) and water (20mL) at room temperature. MeOH (4 mL) was added, and stirring continuedfor 2 h at room temperature at which point LCMS indicated completeconsumption of the ester. Upon concentration in vacuo, the reactionmixture was acidified to pH 3-4 by adding 2N HCl (20 mL). After 20 minstirring, the reaction mixture was cooled to 0° C., a solid precipitatewas collected by filtration, washed with 3-4 ml water, and dried to give1.59 g (64%) of the corresponding acid 38 as a grayish solid. MS:[M+1]=275. To acid 38 (1.59 g, 5.8 mmol) suspended and stirred in DCM(30 ml) was added EDC (5.6 g, 29.2 mmol), benzyl alcohol (2.5 g, 23.2mmol) and DMAP (3.54 g, 29.2 mmol). After 3 days of stirring at roomtemperature, the reaction was concentrated in vacuo. Water (80 mL) wasadded to the slurry, followed by diethyl ether (40 mL), and the mixturewas stirred vigorously for 40 min, at which point the slurry turned intoa precipitate, and was collected by suction filtration. The solid waswashed with water and small amount of diethyl ether, and dried to give1.65 g (78%) benzyl ester 39 as a white solid. MS: [M+1]=365.

Compound 1,2,4-triazole (1.22 g, 17.7 mmol) in CH₃CN (15 mL) at 0° C.was treated with i-Pr₂NEt (3.24 mL, 18.6 mmol), followed by POCl₃ (0.507mL, 5.44 mmol). The solution was stirred at 0° C. for 2 h. Benzyl ester39 (1.65 g, 4.53 mmol) was added in lot and the resulting suspension washeated in an 80° C. oil bath for 18 h. LCMS showed 5-10% starting lactamremained. In a separate flask, 1,2,4-triazole (307 mg, total 4.9 eq) inCH₃CN (3.8 mL) was treated with i-Pr₂NEt (0.82 mL, total 5.1 eq) andPOCl₃ (0.127 ml; total 1.5 eq) at 0° C. for 2 h. The resulting clearsolution was transferred into the above reaction mixture. After 2 hheating at 80° C., the reaction was cooled to room temperature, waterwas added slowly to quench the reaction (10 min). Upon cooling in an icebath, the solids formed were collected by filtration, washed with water(5 ml), and dried to give 1.61 g (86%) product 40 as a lightly yellowsolid. MS: [M+1]=416.

A solution of KO-t-Bu (0.739 g, 6.59 mmol) in DMF (11 mL) was cooled to−50° C. Ethyl isocyanoacetate (0.810 mL, 7.00 mmol) was added drop wise.The mixture was stirred at −50° C. for 1 h. The above triazoleintermediate 40 (1.61 g, 3.87 mmol) was added. The mixture was stirredat −50° C. for 30 min, and slowly warmed to room temperature over 4-5 h.Saturated NH₄Cl aqueous solution (10 mL) was added, followed by EtOAc(10 mL). The mixture was sonicated to breakup solid chunks, then stirredthoroughly for 30 min. The precipitate was collected by filtration,washed with water, Et₂O, and dried to give crude product as a whitesolid. Filtrate was partitioned between water and EtOAc; aqueous layerwas separated and extracted with EtOAc twice; the combined EtOAc layerwas washed with brine and dried over MgSO₄. Filtration and solventremoval gave a solid residue which was combined with the solid obtainedabove for chromatographic purification, using RediSep 24 g silica-gelcolumn and gradient elution with 0.5 to 5% MeOH in DCM, to give 1.78 g(100%) imidazole 41 as a white solid. MS: [M+1]=460. The benzyl ester 41(1.78 g, 3.87 mmol) was subjected to hydrogenolyis (hydrogen balloon) inthe presence of catalytic amount of 10% Pd on charcoal in a solventmixture of THE (40 mL), MeOH (20 mL) and EtOAc (20 mL) for 20 h. LCMSshowed complete disappearance of the starting material. The solidcatalyst was removed by filtration over Celite, and rinsed repeatedlywith ample amount of 30% MeOH in DCM until almost all products wererecovered (TLC monitor). Filtrate containing the product wasconcentrated in vacuo to give 1.22 g (85%) of acid product 42 wasobtained as a yellowish solid. MS: [M+1]=370.

To the acid 42 (1.22 g, 3.30 mmol) suspended and stirred in THE (25 mL)at 0° C. was added borane dimethylsulfide complex (2M THF; 19 mL, 38mmol) dropwise. Ice bath was removed and the reaction mixture wasstirred at room temperature for 16 h. Upon cooling in an ice bath, thereaction was carefully quenched with MeOH (20 mL), and then stirred atroom temperature overnight. Solvents were removed in vacuo. MeOH wasadded and removed in vacuo two more times. ISCO purification (RediSep 24g column) using a gradient of 1 to 8% MeOH in DCM gave 0.625 g (53%) ofalcohol product 43 as a white solid. MS: [M+1]=356.

Diisopropyl azodicarboxylate (48.3 mg, 0.233 mmol) was added drop-wiseinto a stirring solution of alcohol 43 (37.5 mg, 0.106 mmol), phenol(14.9 mg, 0.158 mmol), and Ph₃P (55.6 mg, 0.212 mmol) in anhydrous THE(0.8 mL) at 0° C. Ice bath was removed and stirring continued at roomtemperature for 16 h. LCMS showed complete disappearance of the startingalcohol. The reaction mixture was partitioned between sat. NaHCO₃ andEtOAc. The organic layer was separated and washed with water, brine, anddried over MgSO₄. The desired product was isolated from the reactionmixture by two consecutive preparative TLC (4% MeOH in DCM, andhexanes/EtOAc/MeOH=47.5/47.5/5, v/v/v) to give 5.3 mg (12%) of productwhich is Compound 9 as a white solid. MS: [M+1]=432. ¹H-NMR (500 MHz,CDCl3) δ: 7.77 (s, 1H), 7.63 (d, 1H, J=3.5 Hz), 7.53 (d, 1H, J=9.0 Hz),7.31 (m, 2H), 7.17 (dd, 1H, J=3.0, 8.5 Hz), 7.08 (d, 2H, J=7.0 Hz), 6.99(t, 1H, J=6.5 Hz), 5.30 (s, 2H), 4.40 (q, 2H, J=7.0 Hz), 3.96 (s, 3H),1.38 (t, 3H, J=7.0 Hz).

Example 17: Synthesis of Compound 10

Compound of Example 17 was synthesized in an analogous synthetic routeas that described for Example 16, using 4-fluoro-phenol in the ultimatestep to give Compound 10 (4.9 mg) as a white solid: MS: [M+1]=450.¹H-NMR (500 MHz, CDCl3) δ: 7.76 (s, 1H), 7.64 (d, 1H, J=3.5 Hz), 7.53(d, 1H, J=8.0 Hz), 7.17 (dd, 1H, J=2.5, 8.0 Hz), 7.01 (m, 4H), 5.26 (s,2H), 4.40 (q, 2H, J=7.0 Hz), 3.96 (s, 3H), 1.40 (t, 3H, J=7.0 Hz).

Example 18: Synthesis of Compound 11

Compound of Example 18 was synthesized in an analogous synthetic routeas that described for Example 16, using 3-methoxy-phenol in the ultimatestep to give Compound 11 (6.1 mg) as a white solid: MS: [M+1]=462.¹H-NMR (500 MHz, CDCl3) δ: 7.76 (s, 1H), 7.63 (d, 1H, J=2.5 Hz), 7.53(d, 1H, J=9.0 Hz), 7.15-7.22 (m, 2H), 6.67 (m, 2H), 6.55 (br dd, 1H,J=2.5, 8.0 Hz), 5.28 (s, 2H), 4.39 (q, 2H, J=7.0 Hz), 3.96 (s, 3H), 3.81(s, 3H), 1.39 (t, 3H, J=7.0 Hz).

Example 19: Synthesis of Compound 12

Compound of Example 19 was synthesized in an analogous synthetic routeas that described for Example 16, using 2,4-dimethylphenol in theultimate step to give Compound 12 (3.1 mg) as a white solid: MS:[M+1]=460. ¹H-NMR (500 MHz, CDCl3) δ: 7.76 (s, 1H), 7.65 (d, 1H, J=3.0Hz), 7.53 (d, 1H, J=9.0 Hz), 7.17 (dd, 1H, J=2.5, 8.5 Hz), 6.98 (m, 3H),5.26 (s, 2H), 4.37 (q, 2H, J=7.0 Hz), 3.96 (s, 3H), 2.26 (s, 3H), 2.20(s, 3H), 1.36 (t, 3H, J=7.0 Hz).

Example 20: Synthesis of Compound 107

To a solution of alcohol 43 where X═F (prepared in an identical mannerto example where X═OCH₃) (60 mg, 0.17 mmol) in THE (0.8 mL) was addedphenol (30 mg, 0.32 mmol), triphenylphosphine (84 mg, 0.32 mmol). Thereaction mixture was stirred at room temperature for 15 min. It was thencooled with an ice bath and DIAD (64 μL, 0.32 mmol) in THE (0.2 mL) wasadded slowly. The ice bath was removed and the reaction mixture wasstirred at room temperature for 18 h. LCMS indicated still the presenceof some starting material. Phenol (10 mg), triphenylphosphine (28 mg)and DIAD (21 μL) were added to the reaction mixture and stirred foranother hour. The solvent was evaporated and the crude material waspurified by Chromatography (RediSep 12 g silica-gel column. Elutingsolvent: EtOAc) and prep TLC (eluting solvent: 5% MeOH/47.5% EtOAc/47.5%Hexanes) to give 11.4 mg (yield 16%) of the product Compound 107.[M+1]=421). H¹NMR (CDCl₃) δ 7.92 (1H, dd, J=3.5, 8.5 Hz), 7.80 (1H, s),7.63 (1H, dd, J=5, 10 Hz), 7.38 (1H, m), 7.31 (2H, t, J=8.5 Hz), 7.07(2H, d, J=8.5 Hz), 7.00 (1H, t, J=8.5 Hz), 5.3 (2H, s), 4.39 (2H, q, J=7Hz), 1.38 (3H, t, J=7 Hz).

Example 21: Synthesis of Compound 111

To a suspension of alcohol 43 (X=Me) (160 mg, 0.47 mmol) in acetonitrile(9 mL) was added POBr₃ (405 mg, 1.41 mmol). The reaction mixture washeated at 80 C for 5 h. The reaction mixture was cooled down with an icebath and sat. aq. NaHCO₃ solution was added. The resulting solution wasextracted with DCM (3×). The combined organic phases were washed withbrine and dried over MgSO₄. The solvent was concentrated to afford thedesired product, 166 mg, 88% yield, [M+1]=403).

To a suspension of the above alkyl bromide derivative (30 mg; 0.075mmol) in deoxygenated DME (2.7 mL) was added 3-pyridine boronic acid (14mg, 0.11 mmol) and a 2M Na₂CO₃ solution (0.22 mL, 0.44 mmol). Thesuspension was stirred at room temperature for 5 min, then PdCl₂(PPh₃)₂(10 mg, 0.015 mmol) was added. The suspension was heated in a MW at 85 Cfor 1 hour. The reaction mixture was cooled and diluted with water andextracted with EtOAc (twice). The combined extracts were washed withbrine and dried over MgSO₄. Filtration and concentration gave the crudeproduct which was purified by 2 prep TLC (eluting system: 3% MeOH inDCM) to give 5.3 mg (yield 18%) of the product Compound 111. MS:[M+1]=401. H¹NMR (CDCl₃) δ 8.66 (1H, bs), 8.48 (1H, bs), 7.96 (1H, s),7.79 (1H, s), 7.66 (1H, d, J=8 Hz), 7.50 (1H, d, J=8 Hz), 7.43 (1H, d,J=7 Hz), 7.23 (1H, m), 4.42 (2H, q, J=7 Hz), 4.18 (2H, s), 2.54 (3H, s),1.44 (3H, t, J=7 Hz).

Example 22: Synthesis of Compound 48

To alcohol 43 (186 mg, 0.523 mmol) stirring in DMSO (1 mL) anddichloromethane (2.5 mL) at room temperature was added triethylamine(0.394 mL, 2.82 mmol) and pyridine sulfur trioxide complex (225 mg, 1.41mmol). After 3 h stirring, the reaction was quenched with water (5 mL),and extracted with ethyl acetate three times. The combined organicsolution was washed with water, brine, and dried over MgSO₄. Thealdehyde product 57 was isolated by ISCO flash column chromatography(RediSep 4 g column) using a gradient elution of 0.5 to 8% MeOH in DCM.84.4 mg (46%) was obtained as a yellowish foamy solid. MS: [M+1]=354.

To a stirring solution of aldehyde 57 (15.5 mg, 0.0439 mmol) in1,2-dichloroethane (0.3 mL) at room temperature was added pyrrolidine(5.5 uL, 0.0658 mmol). After 2 min stirring, the solution turned clear,and NaBH(OAc)₃ (14.4 mg) was added. The reaction mixture was stirred for4 h, and was quenched with saturated NaHCO₃, and extracted with ethylacetate three times. The combined organic layer was washed with water,brine, and dried over Na₂SO₄. Prep TLC with 10% MeOH in DCM gave 13.1 mg(73%) of the desired Compound 48 as a clear filmy solid. MS: [M+1]=409.¹H-NMR (500 MHz, CDCl3) δ: 7.74 (s, 1H), 7.62 (d, 1H, J=3.0 Hz), 7.51(d, 1H, J=9.0 Hz), 7.14 (dd, 1H, J=3.5, 9.0 Hz), 4.42 (q, 2H, J=6.5 Hz),3.94 (s, 3H), 3.87 (br s, 2H), 2.65 (br s, 4H), 1.79 (br s, 4H), 1.44(t, 3H, J=7.0 Hz).

Example 23: Synthesis of Compound 49

Compound of Example 23 was synthesized in an analogous synthetic routeas that described for Example 22, using morpholine in the ultimate stepto give the compound of Example 23 as a clear filmy solid: MS:[M+1]=425. ¹H-NMR (500 MHz, CDCl3) δ: 7.75 (s, 1H), 7.63 (d, 1H, J=3.0Hz), 7.52 (d, 1H, J=9.5 Hz), 7.15 (dd, 1H, J=3.0, 9.0 Hz), 4.42 (q, 2H,J=7.5 Hz), 3.95 (s, 3H), 3.76 (br s, 2H), 3.71 (br s, 4H), 2.57 (br s,4H), 1.44 (t, 3H, J=8.0 Hz).

Example 24: Synthesis of Compound 50

Compound of Example 24 was synthesized in an analogous synthetic routeas that described for Example 22, using diethylamine in the ultimatestep to give the compound of Example 24 as a clear filmy solid: MS:[M+1]=411. ¹H-NMR (500 MHz, CDCl3) δ: 7.74 (s, 1H), 7.64 (br d, 1H,J=3.0 Hz), 7.51 (d, 1H, J=9.0 Hz), 7.15 (dd, 1H, J=2.5, 9.0 Hz), 4.43(q, 2H, J=6.5 Hz), 3.96 (s, 3H), 3.86 (br s, 2H), 2.64 (br s, 4H), 1.44(t, 3H, J=8.5 Hz), 1.15 (br s, 6H).

Example 25: Synthesis of Compound 51

Compound of Example 25 was synthesized in an analogous synthetic routeas that described for Example 22, using methyl benzyl amine in theultimate step to give the compound of Example 25 as a clear filmy solid:MS: [M+1]=459. ¹H-NMR (500 MHz, CDCl3) δ: 7.75 (s, 1H), 7.63 (d, 1H,J=3.0 Hz), 7.51 (d, 1H, J=8.5 Hz), 7.36 (br d, 2H, J=8.0 Hz), 7.30 (m,2H), 7.23 (m, 1H), 7.15 (dd, 1H, J=3.0, 9.0 Hz), 4.38 (q, 2H, J=7.5 Hz),3.95 (s, 3H), 3.85 (br s, 2H), 3.63 (br s, 2H), 2.25 (s, 3H), 1.41 (t,3H, J=7.0 Hz).

Example 26: Synthesis of Compound 170

Isobutyramidoxime (41.8 mg, 0.41 mmol) and ester 48 (27.9 mg, 0.0683mmol) in a round bottom flask was azeotroped in toluene on a Rotavapseveral times, suspended in anhydrous THE (0.6 mL), and then cooled to0° C. NaH (60% oil suspension; 10.9 mg, 0.273 mmol) was added. Ice bathwas removed and the reaction mixture was stirred at RT for 20 min beforebeing heated at 70° C. for 6 hrs, and cooled. Water (4 mL) was added,and the mixture was extracted with EtOAc three times. The combinedorganic solution was washed with brine and dried over MgSO4. Prep. TLCwith 10% MeOH in EtOAc gave 10.4 mg (34%) of the desired productCompound 170 as a clear filmy solid. MS: [M+1]=447.

Example 27: Synthesis of Compound 52

The starting alcohol 43 (160 mg, 0.45 mmol) was treated with phosphorousoxide tribromide (400 mg, 1.4 mmol) in acetonitrile (10 ml) at 80° C.for 5 h. The reaction was then cooled down to 0° C., quenched with sat.NaHCO₃, and extracted with dichloromethane twice. Combineddichloromethane solution was washed with brine and dried over MgSO₄.Filtration and solvent removal in vacuo gave 173.3 mg (92%) of thebromide as a yellowish foamy solid. MS: [M+1]=418.

To a suspension of bromide (55 mg, 0.131 mmol) in dimethoxyethane (2 ml;degassed) was added 2M Na₂CO₃ (0.39 ml, 0.78 mmol) and 3-chlorophenylboronic acid (42.2 mg, 0.27 mmol). The reaction mixture was stirred atroom temperature for 2 min, then Pd(PPh₃)₄ (75 mg, 0.065 mmol) wasadded, and the suspension was heated in a 85° C. oil bath for 90 min.Upon cooling, the reaction mixture was diluted with EtOAc and washedwith brine. The aqueous layer was separated and extracted with EtOActhree times. All organic layers were pooled and dried over Na₂SO₄, thenfiltered and solvent was removed in vacuo. The product was isolated bysuccessive prep TLC purifications, using 20% hexanes in EtOAc followedby 5% MeOH in DCM. 9.6 mg product (Compound 52) was obtained as abrownish solid. MS: [M+1]=450. ¹H-NMR (500 MHz, CDCl3) δ: 7.75 (s, 1H),7.64 (d, 1H, J=3.0 Hz), 7.51 (d, 1H, J=9.5 Hz), 7.31 (br s, 1H), 7.23(br s, 1H), 7.17 (m, 3H), 4.43 (q, 2H, J=7.0 Hz), 4.15 (s, 2H), 3.96 (s,3H), 1.44 (t, 3H, J=8.0 Hz).

Example 28: Synthesis of Compound 53

Compound of Example 28 was synthesized in an analogous synthetic routeas that described for Example 27, using 3-cyanophenyl boronic acid inthe ultimate step to give the compound of Example 28 as a brownishsolid: MS: [M+1]=441. ¹H-NMR (500 MHz, CDCl3) δ: 7.75 (s, 1H), 7.66 (brs, 1H), 7.64 (d, 1H, J=3.0 Hz), 7.61 (br d, 1H, J=7.5 Hz), 7.39 (t, 1H,J=7.5 Hz), 7.16 (dd, 1H, J=3.5, 9.5 Hz), 4.45 (q, 2H, J=7.0H), 4.20 (s,2H), 3.96 (s, 3H), 1.45 (t, 3H, J=7.0 Hz).

Example 29: Synthesis of Compound 54

Compound of Example 29 was synthesized in an analogous synthetic routeas that described for Example 27, starting with the alcohol whereR₁=methyl, and using 2-chlorophenyl boronic acid in the ultimate step togive the compound of Example 29 as a brownish solid: MS: [M+1]=434.

Example 30: Synthesis of Compound 101

Compound of Example 30 was synthesized in an analogous synthetic routeas that described for Example 27, starting with the alcohol whereR₁=methyl, and using phenyl boronic acid in the ultimate step to givethe compound of Example 30 as a brownish solid product which waspurified by chromatography (RediSep 4 g silica-gel column. Elutingsolvent: EtOAc) then a prep TLC (eluting system: 40% DCM/40% Hexanes/17%EtOAc/3% MeOH) to give 5.9 mg (yield 31%) of the product Compound 101.MS: [M+1]=402. H¹NMR (CDCl₃) δ 7.96 (1H, s), 7.77 (1H, s), 7.55 (1H, m),7.47 (1H, m), 7.32 (5H, m), 4.41 (2H, q, J=7 Hz), 4.17 (2H, s), 2.53(3H, s), 1.43 (3H, t, J=7 Hz).

Example 31: Synthesis of Compound 102

To a suspension of the bromide in EtOAc (2 mL) and MeOH (2 mL) was addedactivated 10% Pd/C (5 mg). The suspension was stirred under a hydrogenatmosphere for 48 h. The solution was filtered over celite. The filtratewas concentrated and purified by chromatography (RediSep 4 g silica-gelcolumn. Eluting solvent: EtOAc) to give 15.9 mg (33%) of the desiredproduct Compound 102. MS: [M+1]=324. H¹NMR (CDCl₃) δ 7.96 (1H, s), 7.78(1H, s), 7.49 (1H, d, J=9 Hz), 7.42 (1H, d, J=8 Hz), 4.43 (2H, q, J=7.5Hz), 2.53 (3H, s), 2.44 (3H, s), 1.45 (3H, t, J=7.5 Hz).

Example 32: Synthesis of Compound 108

To a suspension of the bromide derivative where R₁═OMe, (18 mg; 0.043mmol) in deoxygenated DME (2 mL) was added 2-chlorophenyl boronic acid(10 mg, 0.065 mmol) and a 2M Na₂CO₃ solution (0.13 mL, 0.26 mmol). Thesuspension was stirred at room temperature for 15 min, then PdCl₂dppf (7mg, 0.009 mmol) was added. The suspension was heated in an oil bath at85 C for 1 hour. The reaction mixture was diluted with water andextracted with EtOAc (twice). The combined extracts were washed withbrine and dried over Na₂SO₄. Filtration and concentration gave the crudeproduct which was purified by PrepTLC (eluting system: 5% MeOH/47.5%Hex/47.5% EtOAc) to give 3.5 mg (yield 18%) of the product Compound 108.MS: [M+1]=451. H¹NMR (CDCl₃) δ 7.77 (1H, s), 7.63 (1H, d, J=3 Hz), 7.52(1H, d, J=11.5 Hz), 7.36 (1H, m), 7.31 (1H, m), 7.18 (2H, m), 7.14 (1H,dd, J=3, 9 Hz), 4.38 (2H, q, J=7 Hz), 4.27 (2H, s), 3.94 (3H, s), 1.41(3H, t, J=7 Hz).

Example 33: Synthesis of Compound 55

To a solution of compound 58 (6.6 g, 33.5 mmol) in dichloromethane (100mL) were added DIPEA (8.65 g, 67 mmol), HOBt (5.4 g, 36.85 mmol) andEDCI (9.6 g, 50.3 mmol). After about 15 min stirring, to the homogeneousreaction mixture was added a solution of 2,4-dimethoxybenzyl amine (5.6g, 33.5 mmol) in dichloromethane (50 mL) dropwise under nitrogenatmosphere. The resulting mixture was stirred under nitrogen atmosphereat room temperature for 16 h. The reaction mixture was washedsuccessively with 1N NaOH (100 mL), water (100 mL) and brine (100 mL).The organic phase was then dried over Na₂SO₄ and evaporated to give acrude solid product 59 that crystallized from ethyl ether. Filtrationand open air suction drying afforded an off-white solid pure product 9.8g (96%), (MS: [M+1]=347).

To a solution of compound 59 (9.8 g, 28.3 mmol) in MeOH/EtOAc (1:1, 100mL) was added 10% wet Pd—C (1.8 g, 10% mmol). After three consecutivevacuuming and flushing with nitrogen, the heterogeneous reaction mixturewas subjected to a balloon hydrogenation at atmosphere pressure up untilthe absorption of hydrogen ceases, about 4 h. The reaction mixture wasfiltered through a celite pad and evaporated to afford the pure desiredproduct 60 as a brown oil 8.63 g (96%), (MS: [M+1=317]). This productwas used directly in the next step.

To a solution of compound 60 (8.63 g, 27.3 mmol) in dichloromethane (100mL) was added triethylamine (5.5 g, 54.6 mmol). The mixture was cooledwith ice bath and treated with bromo acetyl chloride (5.2 g, 32.76 mmol)under nitrogen atmosphere. The ice bath was removed and the mixture leftstirring for 18 h. The reaction mixture was washed successively withsaturated NaHCO₃ (100 mL), water (100 mL) and brine (100 mL). Theorganic phase was then dried over Na₂SO₄ and evaporated to give a crudesolid product 61. The crude product was crystallized from methanol,filtered and dried to afford a brown solid pure product 10.3 g (87%),[MS: 439].

To a solution of compound 61 (10 g, 22.9 mmol) in DMF (1000 mL) wasadded K₂CO₃ (4.8 g, 45.8 mmol). The mixture was heated at 50° C. for 24h. LCMS showed a complete conversion to the desired product. The mixturewas cooled to room temperature and the inorganic solid was filtered. Thesolvent was removed under high vacuum. The resulting crude product 62was crystallized from methanol, filtered and dried to give a pure brownsolid product 6.4 g (78%), (MS: [M+1]=357).

To compound 62 (4.46 g, 12.52 mmol) dissolved in 2.5:1 THF/DMF (50 mL)at −20° C. was added t-BuOK (97%, 1.88 g, 16.28 mmol). The mixture waswarmed to 25° C., and after stirring for 30 min was cooled again to −20°C. Following dropwise addition of diethyl chlorophosphate (2.35 mL,16.28 mmol), the mixture was stirred for 3 h while warming from −20 to25° C. The reaction mixture was re-cooled to 0° C. and to it was addedethyl isocyanoacetate (1.92 mL, 17.53 mmol). Subsequent cooling to −78°C. was followed by addition of t-BuOK (97%, 1.88 g, 16.28 mmol) andstirring at RT for 5 h. Progress was monitored by LC/MS. The reactionwas quenched by addition of 1:1 saturated NaHCO₃/H₂O (140 mL), theprecipitate was filtered, washed with H₂O and air dried overnight toafford 4.81 g (85%) of imidazole derivative 63 as a yellow solid (MS:[M+1]=452).

To compound 63 (4.81 g, 10.65 mmol) in dichloromethane (35 mL) at 0° C.was added trifluoroacetic acid (35 mL) followed by dropwisetrifluoromethanesulfonic acid (1.9 mL, 21.31 mmol). The mixture waswarmed to RT, stirred for 2 h, then concentrated to afford a residuewhich was dissolved in dichloromethane (120 mL). The crude solution waspartitioned between chilled saturated NaHCO₃ and dichloromethane. Theorganic extractions were combined, dried (MgSO₄), filtered andconcentrated to afford 3.2 g (99%) of deprotected product 64 (brownsolid) of sufficient purity to take on the next step (MS: [M+1]=302).

To lactam 64 (51.8 mg, 0.172 mmol) and N,N-dimethyl-p-toluidine (93.0mg, 0.688 mmol) stirring in chlorobenzene (1 ml) under nitrogen wasadded POCl₃ (52.7 mg, 0.344 mmol). The reaction was then heated at 135°C. for 2 h. Upon cooling to room temperature, phenoxy acetic acidhydrazide (228.4 mg, 1.36 mmol) was added in situ to the imino-chloride65, followed by DIPEA (90 ul). The reaction was stirred at roomtemperature for 30 min, then heated at 100° C. for 90 min. The reactionmixture was cooled, saturated NaHCO₃ (aq.) was added, and extracted withethyl acetate three times; combined organic layer was washed with brine,and dried over MgSO₄. After filtration and concentration, the product asCompound 55 was isolated by ISCO flash column chromatography (RediSep 4g column, 1 to 10% MeOH in DCM as eluting gradient) as a white solid,Wt: 8.6 mg. MS: [M+1]=432. ¹H-NMR (500 MHz, CDCl3) δ: 7.81 (s, 1H), 7.71(d, 1H, J=3.5 Hz), 7.52 (d, 1H, J=9.0 Hz), 7.32 (m, 2H), 7.21 (dd, 1H,J=2.5, 8.5 Hz), 7.11 (d, 2H, J=8.5 Hz), 7.02 (m, 1H), 5.44 (s, 2H), 4.38(q, 2H, J=7.5 Hz), 3.94 (s, 3H), 1.39 (t, 3H, J=7.0 Hz).

Example 34: Synthesis of Compound 56

Compound of Example 34 was synthesized in an analogous synthetic routeas that described for Example 33, using 4-fluoro-phenoxy acetic acidhydrazide in the ultimate step to give the compound of Example 34 as ayellowish solid: MS: [M+1]=450. ¹H-NMR (500 MHz, CDCl3) δ: 7.82 (s, 1H),7.73 (d, 1H, J=3.5 Hz), 7.53 (d, 1H, J=10.0 Hz), 7.22 (dd, 1H, J=3.5,9.0 Hz), 7.08-6.99 (m, 4H), 5.41 (s, 2H), 4.41 (q, 2H, J=7.0 Hz), 3.95(s, 3H), 1.42 (t, 3H, J=6.5 Hz).

Example 35: Synthesis of Compound 103

Compound of Example 35 was synthesized in an analogous synthetic routeas that described for Example 33, using 2-methoxy acetic acid hydrazidein the ultimate step to give the compound of Example 35 as a yellowishsolid: MS: [M+1]=370.

Example 36: Synthesis of Compound 118

Acetamide oxime (8.4 mg, 0.108 mmol) was azeotroped in toluene threetimes on a Rotavap, then suspended in THE (1.0 mL). NaH (60% mineralsuspension; 3.3 mg, 0.081 mmol) was added, and the mixture was stirredat RT for 10 min. Ester 55 (23.2 mg, 0.054 mmol) was added next. After40 min stirring at RT, the reaction mixture was heated at 70° C. for 4h. Upon cooling, cold water (5 mL) was added to the reaction mixture,and ppts were collected by filtration, washed with water, and dried togive 9.7 mg (41%) of the desired product as a yellowish solid. MS:[M+1]=442.

Example 37: Synthesis of Compound 128

Compound of Example 37 was synthesized in an analogous synthetic routeas that described for Example 36 above, using ester Compound 103 in theultimate step to give the compound of Example 37 as a brownish solid:MS: [M+1]=380.

Example 38: Synthesis of Compound 130

Compound of Example 38 was synthesized in an analogous synthetic routeas that described for Example 36, starting with ester Compound 103 andcondensing with isobutyramidoxime to give the compound of Example 38 asa yellowish solid: MS: [M+1]=408.

Example 39: Synthesis of Compound 119

To the carboxylic acid (13.9 mg, 0.0345 mmol; obtained through LiOHhydroxysis of the precursor ester 55) stirring in DCM (0.2 mL) was addedNeopentyl alcohol (30.4 mg, 0.345 mmol), DMAP (4.2 mg, 0.0345 mmol), andEDC (20 mg, 0.104 mmol). After five hour stirring, the reaction mixturewas diluted with EtOAc, washed with sat. NH₄Cl, sat. NaHCO₃, brine, anddried over MgSO4. Silica gel chromatographic purification using agradient of 0 to 8% MeOH in EtOAc gave 11.7 mg (72%) of the desiredproduct Compound 119 as a yellowish solid. MS: [M+1]=474.

Example 40: Synthesis of Compound 120

Compound of Example 40 was synthesized in an analogous synthetic routeas that described for Example 39 above, using 2-propyl alcohol in theultimate step to give the compound of Example 40 as a yellowish solid:MS: [M+1]=446.

Example 41: Synthesis of Compound 129

Compound 103 (Scheme 18a) (66.1 mg, 0.179 mmol) was hydrolyzed in asolvent system of THF/water/MeOH (1.8 ml total, 6/5/1 ratio) by treatingwith LiOH (21.4 mg, 0.895 mmol) at RT for 2 h. Dil. HCl was added toacidify (pH˜3) the reaction mixture. The precipitate was collected byfiltration, washed with water, and dried to give 49.0 mg (80%) of theacid as a brownish solid.

The acid thus obtained was stirred in DMF (0.7 mL) at 0° C. NaHCO₃ (48.1mg, 0.572 mmol) was added, followed by N-bromosuccinamide (96.7 mg,0.543 mmol). After overnight stirring, the reaction was diluted withEtOAc, and washed with sat. NaHCO₃. Aq. Layer was separated andextracted with EtOAc. Combined organic layer was washed with brine,dried over MgSO4, filtered, and concentrated. The product bromide wasobtained by silica gel column chromatography with a gradient elution of0 to 13% MeOH in EtOAc as a white solid (Compound 129). Wt: 28.6 mg(53%). MS: [M+1]=377.

Example 42: Synthesis of Compound 131

Compound 129 (22.6 mg, 0.060 mmol) was hydrogenated over 10% Pd—C inEtOAc (1 mL) and MeOH (1 mL) for 16 h. Filtration over Celite, andsolvent removal gave 14.9 mg (84%) of the des-bromo product Compound 131as a lightly yellowish solid. MS: [M+1]=298.

Example 43: Synthesis of Compound 122

The phenoxy analog (Scheme 18a, R₁═OPh) of acid 66 (20.4 mg, 0.0506mmol) was suspended and stirred in DCM (0.5 mL) at RT. Carbonyldiimidazole (16.4 mg, 0.101 mmol) was added. After 2 h stirring, theresulting suspension was cooled to 0° C., and ammonia (30 uL) was addeddropwise. After 20 min stirring, ice bath was removed and the reactionwas allowed to proceed at RT for 1 hr. The reaction was concentrated byremoving DCM in vacuo. Water (3 mL) was added, and precipitate wascollected by filtration, washed with water, and dried to give 16.2 mg ofthe crude primary amide which was used without further purification.

The primary amide (16.2 mg, 0.0402 mmol) was treated with POCl₃ (46.2mg, 0.302 mmol) in 1,4-dioxane (0.5 mL) at 95° C. overnight. Thereaction mixture was then quenched with sat. NaHCO₃ (5 mL), cooled to 0°C., and precipitate collected by suction filtration, washed with water,and dried to give 13.6 mg (88%) of the nitrile as a brownish solid,Compound 122. MS: [M+1]=385.

Example 44: Synthesis of Compound 123

To Acid 66 (15.8 mg, 0.0392 mmol) stirring in THE (0.15 mL) and DCM(0.15 ml) was added N,O-dimethylhydroxylamine HCl (4.6 mg, 0.047 mmol)and N-hydroxylbenzotriazole hydrate (6.0 mg). EDC (11.3 mg, 0.0588 mmol)and triethylamine (11.9 mg, 0.118 mmol) were then added, and thereaction was stirred at RT for 12 hrs, diluted with EtOAc, washed withsat. NH₄Cl, brine, and dried over MgSO₄. Filtration and solvent removalin vacuo gave 14.4 mg (82%) of the Weinreb amide which was used withoutfurther purification.

To the Weinreb amide (14.4 mg, 0.0323 mmol) stirring in THF (0.3 mL) at0° C. was added ethyl magnesium bromide etherate (3M; 0.323 mL). Thereaction was allowed to warm to RT and stirred for 14 hrs., quenchedwith sat. NH₄Cl, extracted with EtOAc three times; combined organiclayer washed with brine and dried over MgSO₄. Filtration and solventremoval gave the crude ketone product which was purified by prep. TLCusing 8% MeOH in EtOAc. Wt: 4.6 mg (34%) of Compound 123. MS: [M+1]=416.

Example 45: Synthesis of Compound 124

Weinreb amide (18.0 mg, 0.0403 mmol) described above was treated withDIBAL (1M THF; 0.363 mL) at −78° C. for 1 hr, then still at −78° C.quenched with Rochelle salt solution (20%) overnight. The aq. solutionwas extracted with EtOAc three times; combined organic layer was washedwith brine, and dried over MgSO₄. Filtration and solvent removal invacuo gave 13.7 mg of the crude aldehyde which was used without furtherpurification.

The crude aldehyde (13.7 mg) in DCM (0.7 mL) at RT was treated withDeoxo-Fluor (54.8 mg, 0.248 mmol) for 16 hrs. The reaction was quenchedwith sat. NaHCO₃ (5 mL) for 20 min, extracted with EtOAc three times;combined organic layer washed with brine, and dried over MgSO₄.Filtration and solvent removal followed by prep. TLC purification using10% MeOH in EtOAc gave 7.5 mg (52%) of the desired difluoride Compound124 as a yellowish solid. MS: [M+1]=410.

Example 46: Synthesis of Compound 142

Weinreb amide (8.8 mg, 0.0197 mmol) from above in THE (0.15 mL) at 0° C.was treated with phenylmagnesium bromide (1M THF; 0.54 mL) for 2.5 hrs,quenched with sat. NH₄Cl, extracted with EtOAc twice; combined organiclayer washed with brine and dried over MgSO₄. Filtration and solventremoval gave the crude ketone which was used without furtherpurification. The ketone in THE (0.5 mL) was treated with NaBH₄ (6 mg)at RT for 2 hrs., then quenched with sat. NH₄Cl, extracted with EtOActhree times; combined organic layer washed with brine, and dried overMgSO₄. Filtration and solvent removal gave the crude alcohol which wasused without further purification. The thus obtained alcohol in DCM (1.4mL) was treated with triethylsilane (86.4 mg, 0.75 mmol) andtrifluoroacetic acid (171.0 mg, 1.5 mmol) at 40° C. overnight, thenconcentrated in vacuo, diluted with EtOAc, washed with sat. NaHCO₃,brine, and dried over MgSO₄. Filtration and solvent removal gave thecrude benzyl product which was purified by silica gel columnchromatography using 0 to 12% MeOH in EtOAc as eluent; 3.6 mg ofCompound 142 was obtained as a yellowish solid. MS: [M+1]=450.

Example 47: Synthesis of Compound 106

To lactam 64 (185.7 mg, 0.616 mmol) in chlorobenzene (5 mL) was addedN,N-dimethyl-p-toluidine (333.3 mg, 2.465 mmol) and phosphorousoxychloride (188.9 mg, 1.232 mmol). The reaction mixture was heated at135° C. for 2 hrs, cooled to RT, and formylhydrazide (296.0 mg, 4.93mmol) was added, followed by diisopropyl ethyl amine (238.8 mg, 1.85mmol). Following 30 min stirring at RT, the reaction was heated at 100°C. for 1 hr., cooled, and sat. NaHCO₃ (15 mL) added, extracted withEtOAc twice; combined organic layer washed with brine, and dried overMgSO4. Filtration and solvent removal gave the crude triazole productwhich was purified by silica gel column chromatography using 0 to 15%MeOH in EtOAc elution, 35.9 mg (18%) was obtained as a brownish solid.MS: [M+1]=326.

The triazole from above in DCM (1 mL) was treated withN-bromosuccinamide (37.6 mg, 0.21 mmol) at 0° C. The reaction wasallowed to warm to RT slowly, and proceeded at RT overnight, dilutedwith EtOAc, washed with sat. NaHCO₃, brine, and dried over MgSO₄.Filtration and solvent removal gave the crude bromide which was purifiedby silica gel column chromatography using 0 to 10% MeOH in EtOAcgradient; 22.9 mg (51%) of Compound 106 was obtained as an off-whitesolid. [MS]: 406.

Example 48: Synthesis of Compound 104

A microwave reaction vessel was charged with phenol (20.3 mg, 0.216mmol), the bromide substrate from Example 47 (29.1 mg, 0.0719 mmol),Cs₂CO₃ (117.0 mg, 0.360 mmol), diethyl 1,3-acetonedicarboxylate (14.5mg, 0.0719 mmol), and DMF (0.5 ml). The vessel was flushed with nitrogengas. CuI (6.8 mg, 0.036 mmol) was added, and the mixture was stirred atRT for 5 min before heated @140° C. under MW radiation conditions for 60min. The reaction mixture was diluted with EtOAc, washed with water; aq.Layer separated and extracted with EtOAc twice; combined organicsolution was washed with brine and dried over MgSO₄. Filtration andsolvent removal gave the crude ether product which was purified by prep.TLC using 5% MeOH in DCM; 6.6 mg of Compound 104 was obtained as ayellowish solid. MS: [M+1]=418.

Example 49: Synthesis of Compound 105

Compound of Example 49 was synthesized in an analogous synthetic routeas that described for Example 48 above, using 3-methoxy phenol in theplace of phenol, to give the compound of Example 49 as a yellowish foamysolid: MS: [M+1]=448.

Example 50: Synthesis of Compound 112

To a solution of Compound 2 (160 mg, 0.49 mmol) in THE (6 mL), water (5mL) and MeOH (1 mL) was added LiOH (59 mg, 2.45 mmol). The solution wasstirred at room temperature for 3 h. The solution was concentrated andthe crude material was acidified with 1N HCl until pH 3-4. No solid wasobserved. EtOAc was added and the organic phase was extracted (3×). Thecombined extracts were washed with brine and dried over MgSO₄.Filtration and concentration gave 112 mg (77% yield) of the desiredcarboxylic acid product as an orange solid MS: [M+1]=300.

To a suspension of acid (30 mg, 0.1 mmol) in dichloroethane (0.2 mL) wasadded thionyl chloride (0.4 mL; 5 mmol) and DMF (20 μL). The resultingsolution was heated at 70 C for 1 hour. Another 0.2 mL of thionylchloride was added and the solution was heated for another 30 min. Thesolvent was removed. The crude material was dried under vacuo.

The crude acid chloride (0.1 mmol) was suspended in isopropanol andstirred at room temperature for 18 h. The solvent was evaporated and thecrude material was purified by chromatography. (RediSep 4 g silica-gelcolumn, eluted with 10% MeOH in DCM) to give 8.6 mg (25% yield) ofproduct Compound 112 [M+1]=342). H¹NMR (CDCl₃) δ 7.90 (1H, d, J=9 Hz),7.79 (1H, bs), 7.63 (1H, bs), 7.36 (1H, bs), 3.48 (1H, m), 2.45 (3H, s),1.43 (6H, d, J=6.5 Hz).

Example 51: Synthesis of Compound 113

The crude acid chloride prepared above (0.066 mmol) was suspended indichloroethane (1 mL) and 2,2-dimethyl-1-propanol (300 mg, 3.4 mmol) wasadded. The solution was stirred at room temperature for 18 h. No productwas formed. To the solution above, was added DMAP (5 mg, 0.004 mmol) andDCC (15 mg, 0.073 mmol). The solution was stirred at room temperaturefor 2 h. The reaction mixture was directly applied on a prep TLC(eluting system: 75 EtOAc in Hexanes) to give 7.2 mg (30% yield) ofproduct Compound 113. MS: [M+1]=370. H¹NMR (CDCl₃) δ 7.91 (1H, dd, J=3,9 Hz), 7.79 (1H, s), 7.61 (1H, dd, J=4.5, 9 Hz), 7.35 (1H, m), 4.11 (2H,s), 2.44 (3H, s), 1.07 (9H, s).

Example 52: Synthesis of Compound 114

The crude acid chloride prepared above (0.066 mmol) was suspended indichloroethane (1 mL) and 2,2,2-trifluoroethanol (0.1 mL, 1.4 mmol)followed by triethylamine (0.6 mL, 4.3 mmol) was added. The solution wasstirred at room temperature for 2 h 30 min. The solvent was evaporatedand the crude material was purified by chromatography. (RediSep 4 gsilica-gel column, eluted with EtOAc) then purified with a prep TLC(eluting system: 70% EtOAc in Hexanes) to give 8.1 mg (32% yield) ofproduct Compound 114 [M+1]=382).

H¹NMR (CDCl₃) δ 7.91 (1H, dd, J=3.5, 9.5 Hz), 7.83 (1H, s), 7.63 (1H,dd, J=4.5, 9.5 Hz), 7.35 (1H, m), 4.77 (2H, m), 2.43 (3H, s).

Example 53: Synthesis of Compound 136

To a solution of acid prepared in Example 50 (100 mg, 0.33 mmol) in DMF(1.5 mL) cooled with an ice bath was added NaHCO₃ (111 mg, 1.32 mmol)followed by NBS (117 mg, 0.66 mmol). The solution was stirred at roomtemperature for 14 h. The reaction mixture was diluted with water andextracted with EtOAc (5×). The combined extracts were washed with brine(2×) and dried over MgSO₄. Filtration and concentration gave a crudeproduct. Chromatography (RediSep 4 g silica-gel column, eluted withEtOAc) to give 93 mg (85% yield) of product Compound 136 [M+1]=334).H¹NMR (CDCl₃) δ 7.87 (1H, dd, J=2.5, 8.5 Hz), 7.72 (1H, s), 7.56 (1H,dd, J=6, 10 Hz), 7.33 (1H, m), 2.44 (3H, s).

Example 54 Synthesis of Compound 139

General coupling procedure: To a solution of Compound 136 (20 mg, 0.061mmol) in degassed DME (0.9 mL) and water (0.1 mL) was added phenylboronic acid (11 mg, 0.092 mmol), cesium carbonate (80 mg, 0.24 mmol)and Pd Cl₂dppf (5 mg, 0.066 mmol). The suspension was heated at 80° C.for one hour. The reaction mixture was diluted with water, extractedwith EtOAc (3×). The combined extracts were washed with brine (2×) anddried over MgSO₄. Filtration and concentration gave a crude productwhich was purified by prep TLC (eluting system: 3% MeOH in EtOAc).

Compound 139 was prepared using phenyl boronic acid. 10.8 mg (54% yield)of product was obtained. MS: [M+1]=332. H¹NMR (CDCl₃) δ 7.87 (1H, dd,J=3.5, 9.5 Hz), 7.85 (1H, s), 7.63 (3H, m), 7.50 (2H, t, J=6.5 Hz), 7.35(2H, m), 2.41 (3H, s).

Example 55: Synthesis of Compound 140

Compound 140 was prepared similarly using 3-pyridine boronic acid. 8.9mg (27% yield) of product was obtained. MS: [M+1]=333. H¹NMR (CDCl₃) δ8.86 (1H, s), 8.63 (1H, d, J=5 Hz), 8.01 (1H, m), 7.90 (2H, m), 7.64(1H, dd, J=5.5, 9 Hz), 7.44 (1H, m), 7.36 (1H, m), 2.39 (3H, s).

Example 56: Synthesis of Compound 152

Compound 152 was prepared using 1-methylpyrazole-4-boronic acid, HCl.12.5 mg (63% yield) of product was obtained. MS: [M+1]=336. H¹NMR(CDCl₃+MeOD₄) δ 9.04 (1H, bs), 7.99 (1H, bs), 7.75 (2H, m), 7.41 (2H,m), 3.95 (3H, s), 2.32 (3H, s).

Example 57: Synthesis of Compound 154

Compound 154 was prepared using 2-methylpyridine-4-boronic acid pinacolester. 7.1 mg (34% yield) of product was obtained. MS: [M+1]=347. H¹NMR(CDCl₃) δ 8.6 (1H, d, J=6 Hz), 7.89 (1H, dd, J=3.5, 8.5 Hz), 7.87 (1H,s), 7.64 (1H, dd, J=5.5, 9 Hz), 7.48 (1H, s), 7.36 (2H, m), 2.64 (3H,s), 2.41 (3H, s).

Example 58: Synthesis of Compound 117

In a 100 mL round-bottom flask, the lactam ester 16′ (2 g, 7.35 mmol;which was prepared in analogous fashion as 16 described in Scheme 11)was dissolved in 60 mL of anhydrous THF. The solution was stirred atroom temperature under a nitrogen atmosphere. LiBH₄ (2 M in THF, 4 mL, 8mmol) was added slowly. The reaction mixture was stirred under anitrogen atmosphere for 18 h. More LiBH₄ (2 M in THF, 2 mL, 4 mmol) wasadded slowly. The reaction mixture was stirred for another 24 h. Amixture of EtOAc/EtOH (20 mL/20 mL) was added to the reaction mixtureand it was concentrated. The residue was taken up in MeOH and silica gelwas added. After volatile solvents were evaporated, the solid was loadedonto a RediSep 40 g silica-gel column. The desired product was elutedwith 5:1 v/v CH₂Cl₂/MeOH. The alcohol was obtained as a white solid(1.14 g, 67% yield). MS: [M+1]=231.

The alcohol (1.14 g, 4.96 mmol) was suspended in 16 mL of HBr 33% inAcOH and heated at 80° C. for 18 h. The solution was cooled down with anice bath and diluted with EtOAc. A white solid could be observed.Slowly, a sat. aq. NaHCO₃ solution was added. Large amount of EtOAc andMeOH were used to solubilize the solid. The organic phase was extracted(3×) and the combined organic phases were washed with brine, dried overMgSO₄. Filtration and concentration gave a crude product which was usedin the next step without further purification. MS: [M+1]=293.

To a solution of alkyl bromide derivative (4.96 mmol) in EtOAc (50 mL),MeOH (200 mL) and THE (50 mL) was added wet 10% Pd/C (250 mg) and theresulting suspension was stirred under a hydrogen atmosphere for 7 days.The suspension was filtered through Celite and the resulting solutionwas concentrated and co-evaporated with toluene. The crude product wasused in the next step without further purification.

To a solution of 1,2,4-triazole (2.7 g, 39.7 mmol) in anhydrous CH₃CN(20 mL) at 0° C. was added i-Pr₂NEt (7.6 mL, 43.6 mmol). Once all thetriazole was dissolved, POCl₃ (1.11 mL, 11.9 mmol) was added. Themixture was stirred at 0° C. for 2 h. The solution was transferred intothe flask containing the lactam (4.96 mmol). The resulting solution washeated in an oil bath at 80° C. for 16 h. The viscous mixture was cooledwith an ice bath and the solvent evaporated. Diluted with EtOAc andwater was added. It was extracted with EtOAc five times. The combinedextracts were washed with brine and dried over MgSO₄. Filtration andconcentration gave a crude product, which was used directly in the nextreaction. MS: [M+1]=266.

A solution of KOtBu (1.11 g, 9.92 mmol) in DMF (10 mL) was cooled to−50° C. under a nitrogen atmosphere. Ethyl isocyanoacetate (1.2 mL, 10.9mmol) was added slowly. The mixture was stirred between −60° C. to −40°C. for 1 h. The above crude 1,2,4-triazolo intermediate from step 4(4.96 mmol) in DMF (5 mL) was added slowly. The mixture was allowed towarm to room temperature over 16 h. Saturated NH₄Cl aqueous solution wasadded and it was extracted with EtOAc three times. The combined extractswere washed with brine (3×) and dried over MgSO₄. Filtration andconcentration gave a crude product. Chromatography (RediSep 24 gsilica-gel column, eluted with 70% EtOAc in Hexanes) to give 296 mg (20%yield for 4 steps) of product. MS: [M+1]=310.

To a solution of ester derivative (260 mg, 0.84 mmol) in THE (6 mL),water (5 mL) and MeOH (1 mL) was added LiOH (117 mg, 4.85 mmol). Thesolution was stirred at room temperature for 3 h. The solution wasconcentrated and the crude material was acidified with 1N HCl until pH3-4. The solid was collected by multiple filtrations to give 178 mg (75%yield) of the desired product. MS: [M+1]=282.

To a suspension of acid (80 mg, 0.28 mmol) in THF (2 mL) was added CDI(50 mg, 0.31 mmol). The suspension was heated at 65 C for 3 h. LCMSindicated that the reaction was incomplete. More CDI (10 mg) was addedand the solution heated for another hour. The solution was cooled downto room temperature and a NH₄OH solution (1 mL) was added. The solutionwas stirred for one hour. The solid was collected by filtration to give33 mg (42%) of the Compound 117 as the desired product as a white solid.MS: [M+1]=281. H¹NMR (MeOD₄) δ 8.1 (1H, s), 7.9 (1H, s), 7.73 (3H, m),7.07 (2H, s), 2.40 (3H, s).

Example 59: Synthesis of Compound 115

To a suspension of Compound 117 (8 mg, 0.029 mmol) and triethylamine (8μL; 0.058 mmol) in THE (1 mL) was added trifluoroacetic anhydride (8 μL;0.058 mmol). The reaction mixture was stirred at room temperature for 16h. LCMS indicated only 30% conversion. More trifluoroacetic anhydride(30 μL) and triethylamine (30 μL) were added. The solution became clearand stirred for another hour. The reaction was quenched with MeOH. Thesolvent was evaporated and the crude material was purified by prep TLC(eluting system: 70% EtOAc in Hexanes) to give 6.6 mg (83%) of theCompound 115. MS: [M+1]=263. H¹NMR (CDCl₃) δ 8.17 (1H, d, J=7 Hz), 7.88(1H, s), 7.67 (3H, m), 2.46 (3H, s).

Example 60: Synthesis of Compound 127

To a suspension of Compound 115 (16 mg, 0.06 mmol) in EtOH (0.8 mL) andwater (0.2 mL) was added hydroxylamine hydrochloride (6 mg, 0.09 mmol)and potassium carbonate (12 mg, 0.09 mmol). The suspension was heated at80° C. for 16 h. The solution was diluted with EtOAc and washed withwater. Aq. Layer was separated and extracted with EtOAc (3×). Thecombined organic phases were washed with brine, dried over MgSO₄.Filtration and concentration gave 12.2 mg (67% yield) of the desiredproduct. MS: [M+1]=296.

A suspension of oxime (10 mg, 0.034 mmol) in acetic anhydride (0.5 mL)was heated at 110 C for 1 hour. Then, the solution was heated at 130 Cfor 1 hour. Finally, the temperature was increased to 140° C. and heatedfor another 2 h. The reaction mixture was cooled down and EtOH (1 mL)was added to the reaction mixture which was heated for 16 h at 80° C.The solvent was evaporated and the crude material was purified by prepTLC (eluting system: EtOAc) to give 6.1 mg (56% yield) of the desiredproduct Compound 127. MS: [M+1]=320). H¹NMR (CDCl₃) δ 8.16 (1H, m), 7.92(1H, s), 7.65 (3H, m), 2.68 (3H, s), 2.46 (3H, s).

Example 61: Synthesis of Compound 133

To a solution of isobutyric acid (19 μL, 0.2 mmol) in THE (0.5 mL) wasadded CDI (10 mg, 0.062 mmol). The solution was stirred at roomtemperature for 2 h. The solution was then transferred into a vialcontaining the oxime derivative described above (12 mg, 0.041 mmol) andheated at 70° C. for 2 h. LCMS indicated that the reaction wasincomplete. Another batch of reagent (isobutyric acid and CDI) wasprepared and added to the reaction mixture which was heated at 70° C.for another hour. LCMS indicated that all starting material wasconsumed. The solvent was evaporated and the crude material wassuspended in isobutyric acid (1 mL) and heated at 130° C. for one hour.The solvent was evaporated and the crude material was purified by PrepTLC (eluting system: 70% EtOAc in Hexanes) to give 6.7 mg (71%) of thedesired product Compound 133. MS: [M+1]=348.

H¹NMR (CDCl₃) δ 8.16 (1H, m), 7.92 (1H, s), 7.65 (3H, m), 3.32 (1H, m),2.46 (3H, s), 1.5 (6H, d, J=7 Hz).

Example 62: Synthesis of Compound 126

Acetamide oxime was azeotroped three times in toluene before use. To asuspension of acetamide oxime (24 mg, 0.32 mmol) in THE (1 mL) was addedNaH 60% in oil dispersion (13 mg, 0.32 mmol). The suspension was stirredat room temperature for 15 min. Compound 3 (50 mg, 0.16 mmol) was added.The vial containing the ester was rinsed with DMF (1 mL) which was addedto the reaction mixture. The resulting brown suspension was stirred atroom temperature for 30 min then heated at 70° C. for 2 h. Thesuspension was quenched with water and the solution was kept in thefridge overnight. The solid was collected by multiple filtrations togive 16 mg (31% yield) of product Compound 126. MS: [M+1]=320. H¹NMR(CDCl₃) δ 8.18 (1H, m), 7.94 (1H, s), 7.67 (3H, m), 2.51 (3H, s), 2.46(3H, s).

Example 63: Synthesis of Compound 125

To a suspension of the carboxylic acid derived from Compound 3 (30 mg,0.11 mmol), N,O-dimethylhydroxylamine hydrochloride (13 mg, 0.13 mmol),1-hydroxybenzotriazole hydrate (17 mg, 0.11 mmol) and triethylamine (46μL, 0.33 mmol) in THE (0.3 mL) and DCM (0.3 mL) was added1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (32 mg, 0.17mmol). The solution was stirred at room temperature for 16 h. Thereaction mixture was quenched with a saturated ammonium chloridesolution and extracted with EtOAc (3×). The combined extracts werewashed with brine and dried over MgSO₄. Filtration and concentrationgave 31.2 mg (88% yield) of an orange solid which was used in the nextstep without further purification. MS: [M+1]=325.

To a solution of above Weinreb amide derivative (31.2 mg, 0.093 mmol) inTHE (0.5 mL) cooled at −78° C. was added a solution of 3 M ethylmagnesium bromide (0.31 mL, 0.93 mmol). The reaction mixture was stirredbelow −10° C. over a period of 60 min. Then, it was quenched with asaturated ammonium chloride solution and extracted with EtOAc (2×). Thecombined extracts were washed with brine and dried over MgSO₄.Filtration and concentration gave a crude product. Chromatography(RediSep 4 g silica-gel column, eluted with 80% EtOAc in Hexanes) togive 11.1 mg (41% yield) of product Compound 125. MS: [M+1]=294. H¹NMR(CDCl₃) δ 8.15 (1H, m), 7.76 (1H, s), 7.65 (3H, m), 3.08 (2H, q, J=7Hz), 2.44 (3H, s), 1.22 (3H, t, J=7 Hz).

Example 64: Synthesis of Compound 132

To a solution of isobutyronitrile (2.6 mL; 29 mmol) in EtOH (30 mL) andwater (10 mL) was added hydroxylamine hydrochloride (2.01 g, 29 mmol)and potassium carbonate (4 g, 29 mmol). The resulting suspension washeated at 80° C. for 16 h. The solvent was removed under vacuo. Theresidue was co-evaporated with toluene. The crude material was washedwith EtOH and filtered to remove the sodium chloride. The filtrate wasevaporated, co-evaporated with toluene several times and dried undervacuo to give 2 g (69%) of N-hydroxybutyramidine.

To a suspension of N-hydroxybutyramidine (47 mg, 0.46 mmol) in THE (1mL) was added NaH 60% in oil dispersion (18 mg, 0.46 mmol). Thesuspension was stirred at room temperature for 30 min. Compound 3 (47mg, 0.15 mmol) in THF (1 mL) was added. The resulting suspension wasstirred at room temperature for 30 min then heated at 70° C. for 2 h.After one hour, only 50% conversion was observed. No change was observedafter another hour. More reagent (N-hydroxybutyramidine and NaH) asdescribed above was prepared and added to the reaction mixture which washeated for another 40 min. At this point, LCMS showed that the reactionwas complete. The suspension was quenched with water. Some MeOH wasadded to help a complete dissolution, and the solution was extractedwith EtOAc (3×). The combined extracts were washed with brine (3×) anddried over MgSO₄. Filtration and concentration gave a crude product.Chromatography (RediSep 4 g silica-gel column, eluted with EtOAc) togive 20 mg (38% yield) of product Compound 132. MS: [M+1]=348. H¹NMR(CDCl₃) δ 8.18 (1H, d, J=8 Hz), 7.93 (1H, s), 7.69 (3H, m), 3.22 (1H,m), 2.46 (3H, s), 1.43 (6H, d, J=9.5 Hz)

Example 65: Synthesis of Compound 161

To a solution of acid derived from Compound 3 (90 mg, 0.32 mmol) in DMF(2 mL) cooled with an ice bath was added NaHCO₃ (108 mg, 1.28 mmol)followed by NBS (114 mg, 0.64 mmol). The solution was stirred at roomtemperature for 18 h. The reaction mixture was diluted with water andextracted with EtOAc (3×). The combined extracts were washed with brine(2×) and dried over MgSO₄. Filtration and concentration gave a crudeproduct. Chromatography (RediSep 4 g silica-gel column, eluted withEtOAc) to give 54 mg (53% yield) of product. MS: [M+1]=316.

To a solution of bromide derivative (30 mg, 0.1 mmol) in dioxane (1 mL)and triethylamine (1 mL) was added TMS-acetylene (71 μL, 0.5 mmol), CuI(2 mg, 0.01 mmol) and PdCl₂(PPh₃)₂ (7 mg, 0.01 mmol). The solution washeated at 110° C. for 6 h. More Pd catalyst (7 mg) and TMS-acetylene(0.2 mL) were added and the reaction mixture heated for an additional 12h. At this time, LCMS showed about 80% conversion. More Pd catalyst (7mg) and TMS-acetylene (0.2 mL) were added and the reaction mixtureheated for an additional 12 h. LCMS showed complete conversion. Thereaction mixture was then diluted with water and extracted with EtOAc(3×). The combined extracts were washed with brine (2×) and dried overMgSO₄. Filtration and concentration gave a crude product. Chromatography(RediSep 4 g silica-gel column, eluted with 70% EtOAc in Hexanes) togive 23 mg (69% yield) of product. MS: [M+1]=334.

To a solution of alkyne derivative (23 mg, 0.069 mmol) in MeOH (0.6 mL)and H₂O (0.2 mL) was added KOH (4 mg, 0.076 mmol) at 0 C. The solutionwas let warm to room temperature over 16 h. The reaction mixture wasdiluted with a saturated aqueous ammonium chloride solution andextracted with EtOAc (2×). The combined extracts were washed with brine(2×) and dried over MgSO₄. Filtration and concentration gave a crudeproduct which was purified by prep TLC (eluting system: 80% EtOAc inHexanes) to give 8.1 mg (45% yield) of product Compound 161. MS:[M+1]=262. H¹NMR (CDCl₃) δ 8.13 (1H, m), 7.76 (1H, s), 7.62 (3H, m),4.09 (2H, bs), 3.28 (1H, s), 2.44 (3H, s).

Example 66: Synthesis of Compound 146

To a solution of 3-amino-2-methylacrolein (65 mg, 0.76 mmol) inanhydrous THF (2 mL) was added NaH 60% in oil dispersion (30 mg, 0.76mmol). The suspension was stirred at room temperature for 15 min.Compound 115 (50 mg, 0.19 mmol) was added and the reaction mixture washeated at 65° C. for 3 h. The reaction mixture was cooled down with anice bath and water was added. The reaction mixture was stored in thefridge overnight. The solid was collected by filtration to give 27.5 mg(44% yield) of a white solid Compound 146. MS: [M+1]=330. H¹NMR (CDCl₃)δ 8.66 (2H, s), 8.15 (1H, m), 7.89 (1H, s), 7.65 (3H, m), 2.44 (3H, s),2.36 (3H, s).

Example 67: Synthesis of Compound 153

To a suspension of acid derived from Compound 3 (30 mg, 0.11 mmol) indichloroethane (0.2 mL) was added thionyl chloride (1 mL; 13.8 mmol) andDMF (20 μL). The resulting solution was heated at 70° C. for 1 hour. Thesolvent was removed. The crude material was dried under vacuo. The crudematerial was suspended in isopropanol (2 mL) and stirred at roomtemperature for 16 h. The solvent was evaporated, co-evaporated withmethanol and the crude material was purified by prep TLC (elutingsystem: EtOAc) to give 7.2 mg (21% yield) of the product Compound 153.MS: [M+1]=324. H¹ NMR (CDCl₃) δ 8.15 (1H, d, J=8 Hz), 7.81 (1H, s), 7.64(3H, m), 5.32 (1H, q, J=7 Hz), 2.45 (3H, s), 1.43 (6H, d, J=7 Hz).

Example 68: Synthesis of Compound 116

An alternate route to the nitrile-substituted imidazole derivatives wasalso implemented. As an example, Compound 116 was prepared fromimino-derivative as shown in Scheme 22. A solution ofisocyanoacetonitrile (206 mg, 3.12 mmol) in DMF (7 mL) was cooled to−50° C. under a nitrogen atmosphere. KOtBu (320 mg, 2.85 mmol) wasadded. The mixture was stirred at −50° C. for 1 h. The imino derivative(prepared in identical fashion to the imino derivative shown above inScheme 21) (350 mg, 1.24 mmol) was added slowly at −50° C. The mixturewas allowed to warm to room temperature over 16 h. Saturated NH₄Claqueous solution was added and it was extracted with EtOAc three times.The combined extracts were washed with brine (3×) and dried over MgSO₄.Filtration and concentration gave a crude product. Chromatography(RediSep 12 g silica-gel column, eluted with 70% EtOAc in Hexanes) togive 230 mg (70% yield) of the product Compound 116. MS: [M+1]=281.H¹NMR (CDCl₃) δ 7.92 (1H, dd, J=3, 8.5 Hz), 7.81 (1H, s), 7.61 (1H, dd,J=4.5, 9 Hz), 7.38 (1H, m), 2.47 (3H, s).

Example 69: Synthesis of Compound 145

To a suspension of cyanide derivative Compound 116 (50 mg, 0.18 mmol) inEtOH (1.6 mL) and water (0.4 mL) was added hydroxylamine hydrochloride(17 mg, 0.24 mmol) and potassium carbonate (28 mg, 0.2 mmol). Thesuspension was heated at 80° C. for 30 min then cooled down to roomtemperature. A solid precipitate was collected by filtration to give37.8 mg (68% yield) of the desired amino oxime product, [M+1]=314.

A suspension of amide oxime (10 mg, 0.032 mmol) in acetic anhydride (0.5mL) was heated at 140 C for 4 h. The reaction mixture was cooled downand EtOH (1 mL) was added to the reaction mixture which was heated for16 h at 80° C. The solvent was evaporated and the crude material waspurified by prep TLC (eluting system: EtOAc) to give 6.6 mg (61% yield)of the desired product Compound 145. MS: [M+1]=338. H¹NMR (CDCl₃) δ 7.91(1H, dd, J=3.5, 8.5 Hz), 7.89 (1H, s), 7.65 (1H, dd, J=5.5, 10 Hz), 7.35(1H, m), 2.69 (3H, s), 2.45 (3H, s).

Example 70: Synthesis of Compound 149

To a solution of isobutyric acid (30 μL, 0.32 mmol) in THE (0.5 mL) wasadded CDI (16 mg, 0.096 mmol). The solution was stirred at roomtemperature for 2 h. The above amide oxime derivative (10 mg, 0.032mmol) was added and the reaction mixture was heated at 70 C for 45 min.The solvent was evaporated and the crude material was suspended inisobutyric acid (1 mL) and heated at 130° C. for 3 h. The solvent wasevaporated and the crude material was purified by Prep TLC (elutingsystem: 80% EtOAc in Hexanes) to give 10.6 mg (91%) of the desiredproduct Compound 149. MS: [M+1]=366. H¹NMR (CDCl₃) δ 7.90 (1H, dd,J=3.5, 9 Hz), 7.89 (1H, s), 7.66 (1H, dd, J=4.5, 8.5 Hz), 7.36 (1H, m),3.32 (1H, q, J=6.5 Hz), 2.46 (3H, s), 1.49 (6H, d, J=8 Hz).

Example 71: Synthesis of Compound 150

A suspension of the above amide oxime (10 mg, 0.032 mmol) intrifluoroacetic anhydride (0.5 mL) was heated under reflux for 10 min.The solvent was evaporated and the crude material was purified by PrepTLC (eluting system: 80% EtOAc in Hexanes) to give 11.8 mg (94%) of thedesired product Compound 150. MS: [M+1]=392. H¹NMR (CDCl₃) δ 7.92 (2H,m), 7.69 (1H, dd, J=5.5, 9.5 Hz), 7.39 (1H, m), 2.45 (3H, s).

Example 72: Synthesis of Compound 151

To a solution of formic acid (12 μL, 0.32 mmol) in THE (0.5 mL) wasadded CDI (16 mg, 0.096 mmol). The solution was stirred at roomtemperature for 2 h. The above amide oxime derivative (10 mg, 0.032mmol) was added and the reaction mixture was heated at 70° C. for 45min. The solvent was evaporated and the crude material was suspended informic acid (1 mL) and heated at 60° C. for 3 h. The solvent wasevaporated and the crude material was purified by Prep TLC (elutingsystem: 80% EtOAc in Hexanes) to give 2.1 mg (20%) of the desiredproduct Compound 151. MS: [M+1]=324. H¹NMR (CDCl₃) δ 8.83 (1H, s), 7.92(1H, dd, J=3.5, 8 Hz), 7.91 (1H, s), 7.65 (1H, dd, J=4.5, 9 Hz), 7.37(1H, m), 2.45 (3H, s).

Example 73: Synthesis of Compound 155

To a solution of propionic acid (22 μL, 0.29 mmol) in THE (0.5 mL) wasadded CDI (14 mg, 0.087 mmol). The solution was stirred at roomtemperature for 1 hour. The above amide oxime derivative (10 mg, 0.032mmol) in THE (0.5 mL) was added and the reaction mixture was heated at70° C. for 90 min. The solvent was evaporated and the crude material wassuspended in propionic acid (1 mL) and heated at 130° C. for 1 h. Thesolvent was evaporated and the crude material was purified by Prep TLC(eluting system: 80% EtOAc in Hexanes) to give 9.4 mg (94%) of thedesired product Compound 155. MS: [M+1]=352. H¹NMR (CDCl₃) δ 7.91 (1H,dd, J=2, 8.5 Hz), 7.88 (1H, s), 7.65 (1H, dd, J=6, 9.5 Hz), 7.36 (1H,m), 3.01 (2H, q, J=8.5 Hz), 2.46 (3H, s), 1.48 (3H, t, J=8.5 Hz).

Example 74: Synthesis of Compound 160

To a solution of pivalic acid (30 mg, 0.29 mmol) in THF (0.5 mL) wasadded CDI (14 mg, 0.087 mmol). The solution was stirred at roomtemperature for 1 hour. The above amide oxime derivative (10 mg, 0.032mmol) in THE (0.5 mL) was added and the reaction mixture was heated at70° C. for 90 min. The solvent was evaporated and the crude material wassuspended in acetic acid (1 mL) and heated under reflux for 3 h. Thesolvent was evaporated and the crude material was purified by Prep TLC(eluting system: 80% EtOAc in Hexanes) to give 7.4 mg (67%) of thedesired product Compound 160. MS: [M+1]=380. H¹NMR (CDCl₃) δ 7.90 (1H,dd, J=2.7, 9 Hz), 7.88 (1H, s), 7.65 (1H, dd, J=4.5, 9 Hz), 7.35 (1H,m), 2.47 (3H, s), 1.53 (9H, s).

Example 75: Synthesis of Compound 143

A solution of KOtBu (40 mg, 0.36 mmol) in DMF (3 mL) was cooled to −50°C. under a nitrogen atmosphere. p-Tolueneslfonylmethyl isocyanide (76mg, 0.39 mmol) was added. The mixture was stirred at −50° C. for 1 h.The imino-derivative from Scheme 22 (50 mg, 0.18 mmol) was added and themixture was allowed to warm to room temperature over 16 h. SaturatedNH₄Cl aqueous solution was added and it was extracted with EtOAc fivetimes. The combined extracts were washed with brine (3×) and dried overMgSO₄. Filtration and concentration gave the crude product.Chromatography (RediSep 4 g silica-gel column, eluted with 70% EtOAc inHexanes) followed by a prep TLC (eluting system: 30% EtOAc in DCM) togive 22.2 mg (30% yield) of a white solid Compound 143. MS: [M+1]=410.

H¹NMR (CDCl₃) δ 7.91 (2H, d, J=8 Hz), 7.87 (1H, dd, J=2.5, 8.5 Hz), 7.74(1H, s), 7.65 (1H, dd, J=5.5, 9 Hz), 7.34 (3H, m), 2.50 (3H, s), 2.42(3H, s).

Example 76: Synthesis of Compound 144

To 3-ethoxymethacrolein (100 mg, 0.88 mmol) was added 7 N ammonia inmethanol (1.3 mL, 8.8 mmol). The solution was stirred at roomtemperature for 16 h. The solvent was evaporated and the crude yellowsolid corresponding to 3-amino-2-methylacrolein was used in the nextstep without further purification.

To a solution of 3-amino-2-methylacrolein (7 mg, 0.087 mmol) inanhydrous THF (1 mL) was added NaH 60% in oil dispersion (6 mg, 0.16mmol). The suspension was stirred at room temperature for 15 min. Thecyanide derivative (22 mg, 0.079 mmol) in THE (1 mL) was added and thereaction mixture was heated at 65° C. for 1 hour. As described above, anew batch of reagents was prepared with 3-amino-2-methylacrolein (20 mg)and NaH (20 mg) in THE (1 mL), and added to the reaction mixture whichwas heated at 65° C. for another hour. LCMS indicated completion of thereaction. The reaction mixture was quenched with methanol. The solventwas evaporated. The crude material was suspended in water and a solidwas collected by filtration to give 5.2 mg (19% yield) of a light redsolid Compound 144. MS: [M+1]=348. H¹NMR (CDCl₃) δ 8.67 (2H, s), 7.90(1H, d, J=9.5 Hz), 7.85 (1H, s), 7.65 (1H, dd, J=4.5, 9 Hz), 7.34 (1H,m), 2.44 (3H, s), 2.36 (3H, s).

Example 77: Synthesis of Compound 121

To a solution of 1,2,4-triazole, (2.03 g, 29.4 mmol) in anhydrous CH₃CN(20 mL) at 0° C. was added i-Pr₂NEt (5.6 mL, 32.4 mmol). Once all thetriazole was dissolved, POCl₃ (0.82 mL, 8.8 mmol) and compound 16′ (1 g,3.68 mmol) were added. The mixture was stirred at 0° C. for 2 h. Theresulting solution was heated in an oil bath at 80° C. for 16 h. Themixture was cooled with an ice bath, diluted with EtOAc, and water wasadded. It was extracted with EtOAc three times. The combined extractswere washed with brine and dried over MgSO₄. Filtration andconcentration gave 1.05 g (88% yield) of an orange solid which was useddirectly in the next step. MS: [M+1]=324.

A solution of KOtBu (696 mg, 6.2 mmol) in DMF (15 mL) was cooled to −50°C. under a nitrogen atmosphere. Ethyl isocyanoacetate (0.75 mL, 6.8mmol) was added slowly. The mixture was stirred at −50° C. for 1 h. Theabove crude product from step 1 (1 g, 3.1 mmol) was added and themixture was allowed to warm to room temperature over 18 h. SaturatedNH₄Cl aqueous solution was added and it was extracted with EtOAc eighttimes. The combined extracts were washed with brine (3×) and dried overMgSO₄. Filtration and concentration gave the crude product.Chromatography (RediSep 24 g silica-gel column, eluted with 70% EtOAc inHexanes) to give 950 mg (83% yield) of product. MS: [M+1]=368.

To a solution of diester (200 mg, 0.54 mmol) in anhydrous THE (4 mL)stirred at room temperature under a nitrogen atmosphere was added LiBH4(2 M in THF, 0.66 mL, 1.3 mmol). The reaction mixture was stirred undera nitrogen atmosphere for 24 h. A mixture of EtOAc/EtOH (3 mL/3 mL) wasadded to the reaction mixture and it was concentrated. The residue wastaken up in MeOH and silica gel was added. After volatile solvents wereevaporated, the solid was loaded onto a RediSep 4 g silica-gel column.The desired product was eluted with 10:1 v/v CH2Cl2/MeOH. The diol wasobtained as a solid (60 mg, 39% yield). MS: [M+1]=284.

The diol (60 mg, 0.21 mmol) was suspended in 5 mL of HBr 33% in AcOH andheated at 80° C. for 18 h. The solution was cooled down with an ice bathand diluted with EtOAc. Slowly, a saturated aqueous NaHCO3 solution wasadded. The solution was extracted with EtOAc (3×), and the combinedorganic phases were washed with brine, dried over MgSO4. Filtration andconcentration gave a crude product which was used in the next stepwithout further purification. MS: [M+1]=408.

To a solution of dialkyl bromide derivative (0.21 mmol) in EtOAc (10 mL)and MeOH (10 mL) was added wet 10% Pd/C (catalytic amount) and theresulting suspension was stirred under a hydrogen atmosphere for 60 h.The suspension was filtered through Celite and the resulting solutionwas concentrated. The crude product was purified by multiple prep TLC(eluting system: 3% MeOH in EtOAc) to give 6.2 mg (12% yield over 2steps) of the desired product Compound 121. MS: [M+1]=252. H¹NMR (CDCl₃)δ 8.09 (1H, m), 7.74 (1H, s), 7.56 (3H, m), 7.90 (2H, m), 2.42 (3H, s),2.29 (3H, s).

Example 78: Synthesis of Compound 135

Compound 135 was synthesized in an analogous manner to Compound 121 asfollows: To a solution of 1,2,4-triazole (952 mg, 13.8 mmol) inanhydrous CH₃CN (20 mL) at 0° C. was added i-Pr₂NEt (2.6 mL, 15.2 mmol).Once all the triazole was dissolved, POCl₃ (0.45 mL, 4.8 mmol) and thelactam ester (1 g, 3.45 mmol) was added. The mixture was stirred at 0°C. for 2 h. The resulting solution was heated in an oil bath at 80° C.for 16 h. The mixture was cooled with an ice bath, diluted with EtOAc,and water was added. It was extracted with EtOAc three times. Thecombined extracts were washed with brine and dried over MgSO₄.Filtration and concentration gave 1.03 g (87% yield) of an orange solidwhich was used directly in the next step. MS: [M+1]=342. A solution ofKOtBu (658 mg, 5.9 mmol) in DMF (15 mL) was cooled to −50° C. under anitrogen atmosphere. Ethyl isocyanoacetate (0.71 mL, 6.5 mmol) was addedslowly. The mixture was stirred at −50° C. for 1 h. The above crudeproduct from step 1 (1 g, 2.9 mmol) was added and the mixture wasallowed to warm to room temperature over 18 h. Saturated NH₄Cl aqueoussolution was added and it was extracted with EtOAc eight times. Thecombined extracts were washed with brine (3×) and dried over MgSO₄.Filtration and concentration gave the crude product. Chromatography(RediSep 24 g silica-gel column, eluted with 70% EtOAc in Hexanes) togive 1.02 g (90% yield) of product. MS: [M+1]=386.

To a solution of diester (600 mg, 1.56 mmol) in anhydrous THE (8 mL)stirred at room temperature under a nitrogen atmosphere was added LiBH₄(2 M in THF, 3.1 mL, 6.24 mmol). The reaction mixture was stirred undera nitrogen atmosphere for 24 h. A mixture of EtOAc/EtOH (10 mL/10 mL)was added to the reaction mixture and it was concentrated. The residuewas taken up in MeOH and silica gel was added. After volatile solventswere evaporated, the solid was loaded onto a RediSep 12 g silica-gelcolumn. The desired product was eluted with 10:1 v/v CH₂Cl₂/MeOH. Thediol was obtained as a solid (187 mg, 40% yield). MS: [M+1]=302.

The diol (80 mg, 0.27 mmol) was suspended in 7 mL of HBr 33% in AcOH andheated at 80° C. for 48 h. The solution was cooled down with an ice bathand diluted with EtOAc. Slowly, a saturated aqueous NaHCO₃ solution wasadded. The solution was extracted (3×) and the combined organic phaseswere washed with brine, dried over MgSO₄. Filtration, concentration andco-evaporation with toluene gave 100 mg (88% yield) of a beige solidwhich was used in the next step without further purification. MS:[M+1]=426.

To a solution of dialkyl bromide derivative (70 mg, 0.16 mmol) in EtOAc(10 mL) and MeOH (10 mL) was added 10% Pd/C (catalytic amount) and theresulting suspension was stirred under a hydrogen atmosphere for 48 h.The suspension was filtered through Celite and the resulting solutionwas concentrated. The crude product was purified by multiple prep TLC(eluting system 1: 75% EtOAc in Hexanes; eluting system 2: 5% MeOH inEtOAc; eluting system 3: EtOAc) to give 4.1 mg (10% yield) of thedesired product Compound 135. MS: [M+1]=270. H¹NMR (CDCl₃) δ 7.84 (1H,dd, J=2.5, 9 Hz), 7.70 (1H, s), 7.54 (1H, dd, J=5, 8 Hz), 7.30 (1H, m),2.42 (3H, s), 2.28 (3H, s).

Example 79: Synthesis of Compound 134

To a suspension of dialkyl bromide derivative described in Scheme 23,R═H, (30 mg, 0.074 mmol) in EtOH (1 mL), and heated at 80° C. was addeda freshly prepared NaOEt 2M solution (75 μL, 0.15 mmol). The solutionwas heated for 10 min. The solvent was evaporated. The crude materialwas suspended in EtOAc and filtered. The filtrate was concentrated andpurified by prep TLC (eluting system: EtOAc) to give 3.1 mg (12% yield)of the desired product Compound 134. MS: [M+1]=340.

Example 80: Synthesis of Compound 137

To a solution of 5-fluoro-2-nitrobenzoic acid (6.6 g, 35.66 mmol) indichloromethane (100 mL) were added DIPEA (9.22 g, 71.3 mmol), HOBt (6.0g, 39.2 mmol) and EDCI (10.2 g, 53.5 mmol). After about 15 min stirring,to the reaction mixture was added a solution of 2,4-dimethoxybenzylamine (5.96 g, 35.66 mmol) in dichloromethane (50 mL) dropwise undernitrogen atmosphere. The resulting mixture was stirred under nitrogenatmosphere at room temperature for 16 h. The reaction mixture was washedsuccessively with 1N HCl (100 mL), sat. NaHCO₃ (100 mL) and brine (100mL). The organic phase was then dried over MgSO₄. Filtration and solventremoval in vacuo afforded a yellowish solid, wt: 9.3 g (78%). MS:[M+1]=335.

To the nitro benzene analog (9.3 g, 27.8 mmol) suspended and stirred ina solvent mixture of HOAc/THF/MeOH/H₂O (25/100/50/25 mL) at RT was addedZn powder. The mixture was heated to 70° C. for 20 hr., cooled, andfiltered. Solid was rinsed with THF, and the combined filtrate wasconcentrated in vacuo. To the resulting slurry was added sat. NaHCO₃slowly and carefully to avoid excessive forming formation until pH reach7 to 8. The mixture was extracted with EtOAc (3×); combined organiclayer washed with brine, and dried over MgSO4. Filtration and solventremoval gave the crude amine product as a dark brown gummy paste, wt:8.7 g.

To a solution of the aniline from above (8.7 g) in dichloromethane (150mL) was added triethylamine (3.37 g, 33.4 mmol). The mixture was cooledwith ice bath and treated with bromo acetyl chloride (4.81 g, 30.6 mmol)under nitrogen atmosphere. The ice bath was removed and the mixture leftstirring for 72 hr. The reaction mixture was concentrated in vacuo, theresulting slurry treated with Et₂O (100 mL) and water (100 mL). Productprecipitate was collected by filtration, and dried to give 5.6 g productas a brown solid. Et₂O layer was separated from aq. Layer and dilutedwith DCM (50 mL), washed with brine, and dried over MgSO₄. Filtrationand solvent removal gave 5.3 g additional product as a foamy brownsolid. Total wt: 11 g (100%).

To a solution of the bromide (11 g) in DMF (550 mL) was added K₂CO₃ (7.1g, 51.7 mmol). The mixture was heated at 50° C. for 48 hrs. The mixturewas cooled to room temperature and the inorganic solid was filtered off.Filtrate was concentrated in vacuo, treated with water/MeOH (60/10 mL),extracted with DCM (3×); combined organic layer was washed with brineand dried over MgSO₄. Filtration and solvent removal followed by silicagel column chromatography using 5 to 50% EtOAc in DCM gave 3.2 g (36%)of the 7-member lactam as a brownish solid. MS: [M+1]=345.

To the lactam (1.32 g, 3.83 mmol) dissolved and stirred in THF (20 mL)and DMF (3 mL) at −20° C. was added t-BuOK (0.645 g, 5.75 mmol). After30 min stirring at −20° C., diethyl chlorophosphate (1.19 mL, 6.89 mmol)was added dropwise, and the mixture was stirred for 3 h while warmingfrom −20 to 20° C. The reaction mixture was cooled to −78° C. and to itwas added ethyl isocyanoacetate (0.791 mL, 6.89 mmol), followed byaddition of t-BuOK (0.645 g, 5.75 mmol) and stirring continued overnightwhile temperature reached to RT. The reaction was quenched withsaturated NH₄Cl, extracted with EtOAc (2×); combined organic solutionwas washed with brine and dried over MgSO₄. Filtration and solventremoval gave a crude product which was purified by silica gel columnchromatography using 15 to 100% EtOAc in DCM, wt: 0.861 g (47%), as abrown solid. MS: [M+1]=440.

To the imidazole ester from above (861 mg) in dichloromethane (5 mL) at0° C. was added trifluoroacetic acid (5 mL) followed bytrifluoromethanesulfonic acid (0.345 mL). The mixture was warmed to RT,stirred for 3 h, then concentrated to afford a residue which wasdissolved in dichloromethane (50 mL). To which was added sat. NaHCO₃ (50mL), followed by 20 min stirring. pH of the top aq. Layer was testedbasic, and was separated, extracted with DCM (3×); combined DCM solutionwashed with brine and dried over MgSO₄. Filtration and solvent removalgave 0.58 g (100%) of the lactam as a yellowish solid. MS: [M+1]=290.

To lactam (209.1 mg, 0.723 mmol) and N,N-dimethyl-p-toluidine (234.7 mg,1.74 mmol) stirring in chlorobenzene (2.5 mL) under nitrogen was addedPOCl₃ (133.0 mg, 0.867 mmol). The reaction was then heated at 135° C.for 2 h. Upon cooling to room temperature, phenoxy acetic acid hydrazide(189.0 mg, 1.08 mmol) was added, followed by DIPEA (0.455 mL). Thereaction was stirred at room temperature for 30 min, then heated at 100°C. for 60 min. The reaction mixture was cooled, saturated NH₄Cl (aq.)was added, and extracted with ethyl acetate three times; combinedorganic layer was washed with brine, and dried over MgSO₄. Afterfiltration and concentration, the product was isolated by ISCO flashcolumn chromatography using 0 to 10% MeOH in EtOAc, wt: 116.7 mg (36%)of Compound 137 as a yellowish filmy solid. MS: [M+1]=420.

Example 81: Synthesis of Compound 156

Ethyl ester Compound 137 (244.2 mg, 0.582 mmol) in a solvent system ofTHF/water/MeOH (6.0 mL total, 6/5/1 ratio) was treated with LiOH (69.7mg, 2.91 mmol) at RT for 4 hrs, concentrated in vacuo, acidified topH-3, and precipitate collected by filtration. After water washing anddrying, 179.3 mg (79%) of the acid was obtained as a yellowish solid.MS: [M+1]=392.

To the acid (10.8 mg, 0.0276 mmol) stirring in DCM (0.1 ml) at RT wasadded EDCI (21.3 mg, 0.11 mmol), DMAP (6.7 mg, 0.0552 mmol) andisopropyl alcohol (13.2 mg, 0.221 mmol). After 12 hrs, the reaction wasdiluted with EtOAc, washed with sat. NaHCO₃; aq. Layer separated andextracted with EtOAc, combined organic layer washed with brine, anddried over MgSO₄. Filtration and prep. TLC purification of theconcentrate using 10% MeOH in EtOAc gave 8.7 mg (73%) of the isopropylester Compound 156 as a yellowish foamy solid. MS: [M+1]=434.

Example 82: Synthesis of Compound 138

Acetamide oxime (10.7 mg, 0.144 mmol) was azeotroped four times intoluene, and added to the ethyl ester Compound 137 (9.5 mg, 0.0226mmol). THE (0.3 mL) was added, followed by NaH 60% oil suspension (4.5mg, 0.112 mmol). The reaction mixture was stirred at RT for 30 min, thenheated at 70° C. for 2 h, cooled to RT, and solvent removed in vacuo,water (1.5 mL) added to quench the reaction, stirred for 20 min, andcooled to 4° C. Precipitate was collected by filtration, washed withwater, and dried to give 5.2 mg (59%) of the oxadiazole product Compound138 as a light yellow solid. MS: [M+1]=430.

Example 83: Synthesis of Compound 141

Compound of Example 83 was synthesized in an analogous synthetic routeas that described for Example 82, using isobutyramidoxime in place ofacetamide oxime to give the compound of Example 83 as a yellowish solid:MS: [M+1]=458.

Example 84: Synthesis of Compound 157

To the acid prepared above in Example 81 (60.2 mg, 0.154 mmol) stirringin DCM (0.7 mL) at RT was added carbonyl diimidazole (49.9 mg, 0.308mmol). The mixture was stirred for 40 min, then cooled to 0° C., andammonia (0.112 ml) added, warmed to RT while stirring continuedovernight. The reaction was concentrated, water (8 mL) added, andstirred well for 30 min. Resulting precipitate was collected byfiltration, washed with water, and dried to give 51.1 mg (85%) of theprimary amide as a brownish solid. MS: [M+1]=391.

The amide (51.1 mg) from above was treated with POCl₃ (200.8 mg, 1.31mmol) in 1,4-dioxane (0.9 mL) at 90° C. for 14 hrs. Upon cooling to RT,the reaction was carefully quenched with sat. NaHCO₃ (5 mL), stirred for20 min. Precipitate was collected by filtration, washed with water, anddried to give 40.9 mg (85%) of nitrile product Compound 157 as abrownish solid. MS: [M+1]=373.

Example 85: Synthesis of Compound 147

To the nitrile (45.8 mg, 0.123 mmol) in a round bottom flask was addedhydroxylamine hydrochloride (14.5 mg, 0.209 mmol), K₂CO₃ (22.3 mg, 0.161mmol), ethanol (0.6 mL), and water (0.15 mL). The reaction mixture washeated at 80° C. for 30 min, cooled down, and concentrated in vacuo. Theresulting slurry was treated with water (1.5 mL), sonicated to helpmixing, and stirred at RT for 1 h before being cooled to 4° C. Theresulting precipitate was collected by filtration, washed with coldwater (1 mL), and dried to give 40.8 mg (82%) of the adduct as anoff-white solid. MS: [M+1]=406.

Isobutyric acid (31.4 mg, 0.582 mmol) was treated with carbonyldiimidazole (28.4 mg, 0.175 mmol) in THE (0.5 mL) for 2 hrs. TheN-hydroxycarboxamide adduct (11.8 mg, 0.0291 mmol) was added, and thereaction was stirred at RT for 30 min. More isobutyric acid (0.5 mL) wasadded and the reaction mixture was heated at 110° C. for 16 h, cooled,sat. NaHCO₃ (8 mL) added, and extracted with EtOAc (3×); combinedorganic layer washed with brine, and dried over MgSO₄. Prep. TLC (5%MeOH in EtOAc) of the concentrated filtrate gave 11.2 mg (84%) of theoxadiazole Compound 147 as a white solid. MS: [M+1]=458.

Example 86: Synthesis of Compound 148

Compound of Example 86 was synthesized in an analogous synthetic routeas that described for Example 85, using acetic acid in place ofisobutyric acid to give the compound of Example 86 as a white solid: MS:[M+1]=430.

Example 87: Synthesis of Compound 158

Compound of Example 87 was synthesized in an analogous synthetic routeas that described for Example 85, using propionic acid in place ofisobutyric acid to give the compound of Example 87 as a white solid: MS:[M+1]=444.

Example 88: Synthesis of Compound 159

Trifluoroacetic anhydride (196.9 mg, 0.938 mmol) was added to theN-hydroxycarboxamide adduct (19.0 mg, 0.0469 mmol) suspended and stirredin THE (0.2 mL) at RT. After 30 min stirring, the reaction was heated to70° C. for 1 h, cooled to RT, and diluted with EtOAc (10 mL), to whichwas added sat. NaHCO₃ and stirred for 30 min. Aq. Layer was separatedand extracted with EtOAc (1×); combined organic layer was washed withbrine, and dried over MgSO₄. Filtration and solvent removal gave a pasteto which was added nBuOH (5 ml) and HOAc (0.5 mL). This was heated at115° C. for 16 h, cooled and concentrated in vacuo, diluted with EtOAc,washed with sat. NaHCO₃, brine, and dried over MgSO₄. Prep. TLC (5% MeOHin EtOAc) of the concentrated filtrate gave 11.5 mg (51%) of the desiredtrifluoromethyl oxadiazole analog Compound 159 as a yellowish solid. MS:[M+1]=484.

Example 89: Synthesis of Compound 162

To lactam 62 (503.4 mg, 1.42 mmol) stirring in THE (2.9 ml) and DMF (0.8mL) at −20° C. was added tBuOK (240.2 mg). After 30 min stirring,diethyl chlorophosphate (377.7 mg, 2.12 mmol) was added dropwise, andthe reaction mixture was slowly warmed to 8° C. in 3 h before beingcooled down to −20° C. 2.26 mL (2.26 mmol) of oxadiazole isocyanate(ref. JMC, 1996, 39, 170; prepared as 1M THE solution) was added. Thereaction mixture was further cooled to −78° C., tBuOK (238.4 mg) wasadded, and the reaction was slowly warmed to RT overnight. Sat. NH₄Cl (5mL) was added and the mixture was extracted with EtOAc (2×), washed withbrine, and dried over MgSO4. Upon filtration and concentration, theproduct was isolated by silica gel column chromatography using agradient elution of 0 to 10% MeOH in EtOAc to give 246.0 mg imidazoleproduct as a yellowish solid. MS: [M+1]=462.

The imidazole (246.0 mg, 0.533 mmol) obtained above was stirred in DCM(3 ml). Trifluoroacetic acid (3 mL) was added, followed bytrifluoromethyl sulfonic acid (160.0 mg, 1.07 mmol). After 3 h stirring,the reaction was diluted with DCM (20 mL), washed with sat. NaHCO₃; aq.Layer was separated and extracted with DCM (2×); combined DCM solutionwas washed with brine, and dried over MgSO₄. Filtration and solventremoval in vacuo gave 208.7 mg of the crude lactam product as ayellowish flaky solid. [M+1]=312.

Phosphorous oxychloride (29.9 mg, 0.195 mmol) was added to a solution ofthe above obtained lactam (22.5 mg, 0.0723 mmol) andN,N-dimethyl-p-toluidine (51.8 mg, 0.383 mmol) stirring in chlorobenzene(0.45 mL) under nitrogen atmosphere. The reaction mixture was heated at135° C. for 3 h, then cooled to RT. Diisopropylethylamine (75.7 mg,0.586 mmol) and phenoxyacetic hydrazide (50.1 mg, 0.302 mmol) was added,and the reaction mixture was heated at 100° C. for 14 h, cooled to RT,and partitioned between sat. NH₄Cl and EtOAc. Aq. Layer was separatedand extracted with EtOAc; combined EtOAc solution was washed with brine,and dried over MgSO₄. Upon filtration and concentration, the productCompound 162 was isolated by silica gel column chromatography using agradient elution of 0 to 10% MeOH in EtOAc as a yellowish solid. Wt:11.8 mg (37%). MS: [M+1]=442.

Example 90: Synthesis of Compound 163

Compound of Example 90 was synthesized in an analogous synthetic routeas that described for Example 89, using 4-fluorophenoxyacetic hydrazidein place of phenoxyacetic hydrazide to give the compound of Example 90as a yellowish solid: MS: [M+1]=460.

Example 91: Synthesis of Compound 164

Compound of Example 91 was synthesized in an analogous synthetic routeas that described for Example 89, using methoxyacetic hydrazide in placeof phenoxyacetic hydrazide to give the compound of Example 91 as ayellowish solid: MS: [M+1]=380.

Example 92: Synthesis of Compound 165

Preparation of benzyloxy acetic hydrazide: carbonyl diimidazole (1.52 g,9.39 mmol) was added to benzyloxy acetic acid (1.2 g, 7.22 mmol)stirring in THF (60 mL) at 0° C. Ice bath was removed and the stirringcontinued for 1 hr. The resulting cloudy solution was added to hydrazine(0.927 g, 28.9 mmol) stirring in THE (40 mL) at RT. After 16 hrs, thereaction mixture was concentrated to a slurry, to which was added water(120 mL), extracted with DCM (3×); combined DCM solution washed withbrine, and dried over MgSO₄. Filtration and solvent removal gave 0.908 g(70%) of the hydrazide as a clear viscous oil. This was azeotroped intoluene a few times before use. Compound of Example 92 was synthesizedin an analogous synthetic route as that described for Example 89, usingbenzyloxy acetic hydrazide in place of phenoxyacetic hydrazide to givethe compound of Example 92 as a yellowish solid: MS: [M+1]=456.

Example 93: Synthesis of Compound 166

Compound 165 from above (58.5 mg, 0.128 mmol) was treated with 10% Pd—C(catalytic) in EtOAc (4 mL) and MeOH (4 mL) under hydrogen atmospherefor 2 h. Catalyst was removed by filtration over Celite. To the filtratewas added conc. HCl (0.89 mL), and the mixture was stirred at RT for 16h. Excess Na₂CO₃ (aq.) was added, and the solution was extracted withEtOAc (2×); combined organic solution was washed with brine, and driedover MgSO₄. Prep. TLC of the concentrated filtrate using 15% MeOH inEtOAc gave 14.9 mg of the primary amide ([M+1]=417) as a yellowishsolid. This primary amide was treated with phosphorous oxychloride (54.9mg, 0.358 mmol) in 1,4-dioxane (1 mL) at 90° C. for 14 h. Upon cooling,the reaction mixture was diluted with EtOAc, washed with sat. NaHCO₃;aq. layer separated and extracted with EtOAc (1×), combined organicsolution was washed with brine, and dried over MgSO₄. Prep. TLC of theconcentrated filtrate using 5% MeOH in EtOAc gave 5.2 mg of the desirednitrile product Compound 166 as white needles. [M+1]=399.

Example 94: Synthesis of Compound 169

To lactam 62 (2.23 g, 6.24 mmol) stirring in THF (10 mL) and DMF (3 mL)at −20° C. was added tBuOK (1.05 g, 9.36 mmol). After 30 min stirring,diethyl chlorophosphate (1.66 g, 9.36 mmol) was added dropwise, and thereaction mixture was slowly warmed to 8-10° C. in 3 h before beingcooled down to −20° C. 10.0 ml (10.0 mmol) of oxadiazole isocyanate(ref. JMC, 1996, 39, 170; prepared as 1M THE solution) was added. Thereaction mixture was further cooled to −78° C., tBuOK (1.05 g, 9.36mmol) was added, and the reaction was slowly warmed to RT overnight.Sat. NH₄Cl (20 mL) was added and the mixture was extracted with EtOAc(3×), washed with brine, and dried over MgSO₄. Upon filtration andconcentration, the product was isolated by silica gel columnchromatography using a gradient elution of 10 to 100% EtOAc in DCM togive 1.07 g (35%) imidazole product as a yellowish foamy solid. MS:[M+1]=490.

The imidazole (1.07 g, 2.18 mmol) obtained above was stirred in DCM (11mL). Trifluoroacetic acid (11 mL) was added, followed by trifluoromethylsulfonic acid (0.656 g, 4.37 mmol). After 4 h stirring, the reaction wasconcentrated in vacuo, diluted with DCM (50 mL), washed with sat.NaHCO₃; aq. Layer was separated and extracted with DCM (2×); combinedDCM solution was washed with brine, and dried over MgSO₄. Filtration andsolvent removal in vacuo gave 0.872 g of the crude lactam product as abrownish solid. [M+1]=340.

Phosphorous oxychloride (51.0 mg, 0.333 mmol) was added to a solution ofthe above obtained lactam (45.0 mg, 0.133 mmol) andN,N-dimethyl-p-toluidine (89.6 mg, 0.663 mmol) stirring in chlorobenzene(0.60 mL) under nitrogen atmosphere. The reaction mixture was heated at135° C. for 3 h, then cooled to RT. Diisopropylethylamine (137.5 mg,1.06 mmol) and methoxyacetic hydrazide (83.1 mg, 0.798 mmol) was added,and the reaction mixture was heated at 100° C. for 4 h, cooled to RT,diluted with EtOAc, washed with sat.NaHCO₃, brine, and dried over MgSO₄.Upon filtration and concentration, the product Compound 169 was isolatedby silica gel column chromatography using a gradient elution of 0 to 13%MeOH in EtOAc as a brownish solid. Wt: 14.3 mg (26%). MS: [M+1]=408.

Example 95: Synthesis of Compound 171

Compound of Example 95 was synthesized in an analogous synthetic routeas that described for Example 94, using phenoxyacetic hydrazide in placeof methoxyacetic hydrazide to give the compound of Example 95 as ayellowish solid: MS: [M+1]=470.

Example 96: Synthesis of Compound 172

Compound of Example 96 was synthesized in an analogous synthetic routeas that described for Example 94, using 4-fluoro-phenoxyacetic hydrazidein place of methoxyacetic hydrazide to give the compound of Example 96as a yellowish solid: MS: [M+1]=488.

Example 97: Synthesis of Compound 173

Compound of Example 97 was synthesized in an analogous synthetic routeas that described for Example 94, using ethoxyacetic hydrazide in placeof methoxyacetic hydrazide to give the compound of Example 97 as ayellowish solid: MS: [M+1]=422.

Example 98: Synthesis of Compound 174

Compound of Example 98 was synthesized in an analogous synthetic routeas that described for Example 94, using 2-fluoro-phenoxyacetic hydrazidein place of methoxyacetic hydrazide to give the compound of Example 98as a yellowish solid: MS: [M+1]=488.

Example 99: Synthesis of Compound 175

Compound of Example 99 was synthesized in an analogous synthetic routeas that described for Example 94, using 2-chloro-phenoxyacetic hydrazidein place of methoxyacetic hydrazide to give the compound of Example 99as a yellowish solid: MS: [M+1]=504.

Example 100: Synthesis of Compound 176

Preparation of 3-pyridyloxy acetic hydrazide: a solution of ethyl3-pyridyloxy acetate (0.50 g, 2.76 mmol) and hydrazine (0.31 g, 9.66mmol) in isopropyl alcohol (35 mL) was heated at 85° C. for 30 hr.,cooled, and concentrated in vacuo. The resulting white solid wasdissolved in small amount of sat. NaCl solution, and extracted withEtOAc repeatedly. The combined organic solution was dried over MgSO₄.Filtration and solvent removal gave 177 mg of the desired acetichydrazide as a white solid. Residual water moisture was removed byazeotroping in toluene. Compound of Example 100 was synthesized in ananalogous synthetic route as that described for Example 94, using3-pyridyloxy acetic hydrazide in place of methoxyacetic hydrazide togive the compound of Example 100 as a yellowish solid: MS: [M+1]=471.

Example 101: Synthesis of Compound 177

Compound of Example 101 was synthesized in an analogous synthetic routeas that described for Example 94, using 1-naphthoxy acetic hydrazide inplace of methoxyacetic hydrazide to give the compound of Example 101 asan off white solid: MS: [M+1]=520.

Example 102: Synthesis of Compound 179

Compound of Example 102 was synthesized in an analogous synthetic routeas that described for Example 94, using 3-fluorophenoxy acetic hydrazidein place of methoxyacetic hydrazide to give the compound of Example 102as a yellowish solid: MS: [M+1]=488.

Example 103: Synthesis of Compound 178

Phosphorous oxychloride (64.8 mg, 0.422 mmol) was added to a solution ofthe oxadiazolyl imidazole lactam (57.5 mg, 0.169 mmol) andN,N-dimethyl-p-toluidine (114.6 mg, 0.847 mmol) stirring inchlorobenzene (0.70 ml) under nitrogen atmosphere. The reaction mixturewas heated at 135° C. for 3 h, then cooled to RT. Diisopropylethylamine(174.7 mg, 1.35 mmol), t-BuOH (0.3 ml), and 2-hydroxy acetic hydrazide(91.3 mg, 1.01 mmol) was added. The reaction mixture was stirred at RTfor 20 min, then warmed at 50° C. for one hour followed by 80° C.heating for one hour before finally heated at 100° C. overnight. Uponcooling to RT, the reaction was diluted with EtOAc, washed with brine,and dried over MgSO₄. Silica gel column chromatography of theconcentrated filtrate using a gradient elution of 0 to 20% MeOH in EtOAcgave the desired hydroxymethyl triazole product as a yellowish solid.Wt: 18.1 mg (27%). MS: [M+1]=394.

To a solution of hydroxymethyl triazole from above (18.1 mg, 0.046mmol), cyclopentyl bromide (274.0 mg, 1.84 mmol), and HMPA (16.5 mg,0.092 mmol) stirring in THE (0.5 ml) was added NaH (60% suspension; 18.4mg, 0.46 mmol). After 10 min, the reaction was heated at 100° C. for 6hrs, cooled, quenched with sat. NaHCO₃, and extracted with EtOAc (2×),washed with brine, and dried over MgSO₄. Prep. TLC of the concentratedfiltrate using 8% MeOH in EtOAc gave 5.5 mg (26%) of the desired etherCompound 178 as a yellowish solid. [M+1]=462.

Example 104: Synthesis of Compound 168

To a suspension of benzyl glycinate hydrochloride (5 g, 24.8 mmol) inDCM (100 mL) was added EDC.HCl (6.2 g, 33.2 mmol) and triethylamine (5.2mL, 37.2 mmol). The suspension was cooled down to −50° C. then formicacid (1.4 mL, 37.2 mmol) in DCM (5 mL) was added. The reaction mixturewas stirred at −50 C for one hour then at 4° C. for 3 h. The solutionwas diluted with 1N HCl and extracted with DCM (2×). The combinedorganic phases were washed with brine and dried over MgSO4. Filtrationand concentration gave 3.89 g (81% yield) of formylated glycine as anoil (M+1=194)

To a solution of formylated glycine derivative (1 g, 5.2 mmol) in DCM(30 mL) was added triethylamine (3.2 mL, 23 mmol). The solution wascooled down to −50° C. and POCl₃ (1.9 mL, 20.8 mmol) was added slowly.The solution was stirred at −50 C for 10 min, then stirred at roomtemperature for 40 min. The solution turned light red-brown. It wasdiluted with DCM and a 20% sodium carbonate solution (100 mL) was added.The reaction mixture was stirred vigorously for 15 min. The organicphase was separated twice and dried over MgSO4. Filtration andconcentration to give the desired benzyl isocyanoacetate in quantitativeyield which was used in the next step without further purification.

To a solution of 1,2,4-triazole (914 mg, 13.2 mmol) in anhydrous CH₃CN(20 mL) at 0° C. was added i-Pr₂NEt (2.5 mL, 14.6 mmol). Once all thetriazole was dissolved, POCl₃ (0.43 mL, 4.6 mmol) was added. The mixturewas stirred at 0° C. for 2 h. The lactam ester 16′ (1 g, 3.31 mmol) wasadded. The resulting solution was heated in an oil bath at 80° C. for 16h. The mixture was cooled with an ice bath. Diluted with EtOAc thenwater was added. Aq. layer was separated and extracted with EtOAc fourtimes. The combined organic extracts was washed with brine and driedover MgSO₄. Filtration and concentration gave a light yellow solid whichwas used directly in the next step (M+1=354).

A solution of benzyl isocyanoacetate (892 mg, 5.1 mmol) in DMF (10 mL)was cooled to −50° C. under a nitrogen atmosphere. KOtBu (514 mg, 4.6mmol) was added. The mixture was stirred at −50° C. for 1 h. Thetriazole derivative prepared above (900 mg, 2.55 mmol) in DMF (5 mL) wasadded slowly at −50° C. The mixture was allowed to warm to roomtemperature over 16 h. Saturated aqueous NH₄Cl solution was added and itwas extracted with EtOAc three times. The combined extracts were washedwith brine (3×) and dried over MgSO₄. Filtration and concentration gavea crude product. Chromatography (RediSep 24 g silica-gel column, elutedwith 70% EtOAc in Hexanes) to give 886 mg (76% yield) of product(M+1=460).

To a solution of benzyl ester derivative (770 mg, 1.68 mmol) in EtOAc(10 mL) and MeOH (30 mL) was added wet Pd/C (60 mg) and the resultingsuspension was stirred under a hydrogen atmosphere for 48 h. Thesuspension was filtered through Celite and the resulting solution wasconcentrated. The crude debenzylated product (530 mg, 86% yield) wasused in the next step without further purification (M+1=370).

To a suspension of acid (530 mg, 1.44 mmol) in DCM (10 mL) was added CDI(931 mg, 5.75 mmol). The solution was stirred at room temperature for 2h. The solution was cooled down with an ice bath and a NH₄OH solution (6mL) was added. The solution was stirred for 30 min and it wasconcentrated. The solid was collected by filtration and washed withwater to give 422 mg (80%) of the desired product as a brown solid.(M+1=369).

To a suspension of primary amide derivative (422 mg, 1.15 mmol) indioxane (10 mL) was added POCl₃ (160 μL, 1.7 mmol). The suspension washeated at 90° C. for 2 h. The resulting solution was cooled down with anice bath and quenched with a saturated aqueous NaHCO₃ solution. Thesolid was collected by filtration to give 308 mg (77% yield) of thedesired cyanide derivative. (M+1=351).

To a suspension of cyanide derivative (150 mg, 0.44 mmol) in EtOH (4 mL)and water (1 mL) was added hydroxylamine hydrochloride (40 mg, 0.57mmol) and potassium carbonate (67 mg, 0.48 mmol). The suspension wasstirred at room temperature for 16 h. LCMS indicated about 50%conversion. More hydroxylamine hydrochloride (40 mg, 0.57 mmol) andpotassium carbonate (67 mg, 0.48 mmol) were added, and stirred foranother 24 h. The solution was diluted with EtOAc and washed with water.The combined organic phases were washed with brine, dried over MgSO₄.Filtration and concentration gave 145 mg (86% yield) of the desiredproduct. (M+1=384).

To a solution of acetic acid (0.22 mL, 3.8 mmol) in THE (5 mL) was addedCDI (123 mg, 0.76 mmol). The solution was stirred at room temperaturefor 2 h. The solution was then poured into a flask containing the oximederivative (145 mg, 0.38 mmol) and heated at 70 C for 1 hour. Thesolvent was evaporated and the crude material was suspended in aceticacid (8 mL) and heated at 130° C. for one hour. The solvent wasevaporated and the crude material was triturated with water to give 134mg (86%) of the desired product (M+1=408).

To a suspension of ester derivative (50 mg, 0.12 mmol) in THE (1 mL) wasadded lithium aluminum hydride (7 mg, 0.18 mmol). The suspension wasstirred at room temperature for 2 h. LCMS indicated about 70% conversionalong some other side products and some remaining starting material.More lithium aluminum hydride (4 mg) was added and the reaction mixturewas stirred at room temperature for another 30 min. The reaction mixturewas quenched with 1N HCl. The solution was extracted with EtOAc (3×).The combined organic phases were washed with brine, dried over MgSO₄.Filtration and concentration gave 20 mg (45% yield) of the desiredalcohol product. (M+1=366).

To a suspension of alcohol (20 mg, 0.055 mmol) in dioxane (1 mL) wasadded POBr₃ (31 mg, 0.11 mmol). The reaction mixture was heated at 110°C. for 1 hour. The reaction mixture was cooled down with an ice bath andsat. aq. NaHCO₃ solution was added. The resulting solution was extractedwith EtOAc (3×). The combined organic phases were washed with brine anddried over MgSO₄. The solvent was concentrated to give 22 mg (96% yield)of the desired product (M+1=428).

To a vial containing alkyl bromide derivative (22 mg, 0.052 mmol) wasadded 3-fluorophenol (58 mg, 0.52 mmol) in dioxane (1 mL) and potassiumcarbonate (72 mg, 0.52 mmol). The reaction mixture was heated at 90° C.for 1 hour. The reaction mixture was diluted with sat. aq. NaHCO₃solution. The resulting solution was extracted with EtOac (3×). Thecombined organic phases were washed with brine and dried over MgSO₄.Filtration and concentration gave a crude product. Purification by prepTLC (eluting system: EtOAc) to give 5 mg (21% yield) of the desiredproduct Compound 168 (M+1=460). H¹NMR (CDCl₃) δ 7.87 (1H, s), 7.65 (1H,d, J=3.5 Hz), 7.57 (1H, d, J=10 Hz), 7.24 (1H, m), 7.19 (1H, dd, J=3.5,9 Hz), 6.77 (1H, dd, J=2.5, 9.5 Hz), 6.72 (2H, m), 5.26 (2H, s), 3.97(3H, s), 2.48 (3H, s).

Synthesis of Compounds 215-313

Synthesis of Intermediate A (ethyl15-chloro-9-(methoxymethyl)-2,4,8,10,11-pentaazatetracyclo[1.4.0.0²,⁶.0⁸,¹²]heptadeca-1(17),3,5,9,11,13,15-heptaene-5-carboxylate)

Ethyl bromoacetate (Scheme 28) (10.0 gm, 59.87 mmol) solution in 20.0 mLof anhydrous THE was added dropwise to a solution of(2,4-dimethoxybenzyl)amine (10.0 gm, 59.81 mmol) and triethyl amine(6.06 gm, 59.87 mmol) in anhydrous THE (20.0 mL) at 0° C. under nitrogenatmosphere. The reaction mixture was warmed to room temperature andstirred overnight. Brine was added ˜100 mL, and the reaction mixture wasextracted with ethyl acetate (2×˜100 mL). Combined extracts were driedover anhydrous MgSO₄ and concentrated under reduced pressure. Thepurification was performed using combiFlash chromatography, Gradient:20:80 to 50:50 v/v Ethylacetate:Hexane. 7.6 gm (yield 50.2%) of thealkylation product was obtained as a colorless liquid. m/z calculatedfor C₁₃H₁₉NO₄ [M+H]⁺: 254; Obtained: 254.1. The ester (7.5 gm, 29.6mmol) was dissolved in 40.0 mL of methanol. The reaction mixture wascooled and 2N aq. NaOH (88.82 mmol, 44.0 mL) solution was addeddropwise. The reaction mixture was warmed to room temperature andstirred for 2 h. The reaction mixture was diluted with ˜75.0 mL ofwater, cooled in ice bath and neutralized down to ˜5.0 to 4.5 pH using2N aq. HCl. The excess water was concentrated under reduced pressure andair streamed to obtain white solid powder. The solid was dissolved in85:15 v/v, DCM:MeOH (100.0 mL) and filtered, the filtrate was evaporatedto obtain 7.1 gm of carboxylic acid as a white powder (Hygroscopic). m/zcalculated for C₁₁H₁₅NO₄ [M+Na]⁺: 248; Obtained: 248.1.

The above compound (7.0 gm, 31.08 mmol) and 6.14 gm, 31.08 mmol of5-chloroisatoic anhydride were mixed in 70.0 mL of p-Xylene and refluxedat 140° C. temperature for 3 h. The reaction mixture filtered and crudeproduct recrystallized from methanol. 8.5 gm of7-chloro-4-[(2,4-dimethoxyphenyl)methyl]-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2,5-dionewas obtained as a white powder (75.8% yield). m/z calculated forC₁₈H₁₇ClN₂O₄[M+H]⁺: 361; Obtained: 361.1.

The above benzodiazepine-2,5-dione (4 gm, 11.1 mmol) was dissolved inTHF/DMF (57.2/12.7 mL) and cooled at −20° C. temperature. Finely dividedpotassium-tert-butoxide powder (1.9 gm, 16.6 mmol) was added andreaction mixture stirred at −20° C. for 20.0 min. 3.1 gm, 17.7 mmol ofdiethylchlorophosphate was dropwise added to the reaction mixture at−20° C. and allowed to 0-5° C. for 3 h. The reaction mixture was stirredat ambient temperature for 10.0 min. 2.1 gm, 18.4 mmol ofethylisocyanoacetate was added to the reaction mixture at −20° C. andthe reaction mixture was further cooled down to −78° C. 1.9 gm, 16.6mmol of finely divided potassium-tert-butoxide powder was added at −78°C. and the reaction mixture was stirred overnight by slowly warming toambient temperature. The reaction mixture was quenched with saturatedaq. NH₄Cl solution (10 mL), extracted with ethyl acetate (3×20 mL).Combined extracts were dried over anhydrous MgSO₄ and concentrated underreduced pressure. The crude product was recrystallized from ethylacetateto obtain 2.2 gm of ethyl12-chloro-8-[(2,4-dimethoxyphenyl)methyl]-9-oxo-2,4,8-triazatricyclo[8.4.0.0²,⁶]tetradeca-1(14), 3,5,10,12-pentaene-5-carboxylate as a white solid. A second cropwas obtained from the mother liquor to afford another 3.5 g of product(64% yield).

The dimethoxybenzyl protecting group was removed by dissolving the abovecompound (2.2 gm, 4.83 mmol) in DCM (25.0 mL), followed by addition of25.0 mL of trifluoroacetic acid and 1.45 gm, 9.65 mmol oftrifluoromethanesulfonic acid. The reaction mixture was stirred at roomtemperature for 90 min. The reaction mixture was neutralized with aq.NaHCO₃ and the ppts were filtered, washed with water and dried to afford1.9 gm of ethyl12-chloro-9-oxo-2,4,8-triazatricyclo[8.4.0.0²,⁶]tetradeca-1 (14),3,5,10,12-pentaene-5-carboxylate as a solid product. m/z calculated forC₁₄H₁₂ClN₃O₃[M+H]⁺: 306; Obtained: 306.1.

In the first step, the ethyl12-chloro-9-oxo-2,4,8-triazatricyclo[8.4.0.0²,⁶]tetradeca-1 (14),3,5,10,12-pentaene-5-carboxylate from above (1.9 gm, 6.21 mmol) wasdissolved in 25.0 mL of chlorobenzene, followed by addition of 2.52 gm,18.64 mmol of 4,N,N-trimethylaniline, 1.42 gm, 9.32 mmol of POCl₃ andthe reaction mixture was refluxed at 135° C. for 2 h. LCMS shows ˜50%starting material remained unreacted. 1.68 gm, 12.42 mmol of additional4,N,N-trimethylaniline and 0.95 gm, 6.21 mmol of POCl₃ were furtheradded to the reaction mixture at room temperature and refluxed at 135°C. for 1 h. LCMS shows ˜10% starting material remained unreacted. Anadditional 0.84 gm, 6.21 mmol of 4,N,N-trimethylaniline (total 6.0 eq.)and 0.48 gm, 3.11 mmol of POCl₃ (total 3 eq.) were further added to thereaction mixture at room temperature and refluxed at 135° C. for 1 h.

In the second step, 4.67 gm, 44.75 mmol of methoxyaceticacid hydrazide(total 7.2 eq.), followed by 7.71 gm, 59.66 mmol ofN,N-diisopropylethylamine were added to the reaction mixture at roomtemperature and refluxed at 100° C. for 1 h. The reaction mixture wascooled to room temperature and neutralized with aq. NaHCO₃ solution(˜25.0 mL). The organic was extracted with ethyl acetate (75 mL×3),followed by DCM (50.0 mL×3) and washed with brine. The EtOAc organiclayer was separated by filtering the insoluble ppts (0.805 gm pureproduct) and combined organic layers were dried over anhydrous MgSO₄,concentrated under reduced pressure. The crude product was purified byCombiflash chromatography (Mobile phase: 0-10% MeOH:EtOAc) to yield anadditional 0.8 gm of yellow solid. Total yield for the last two steps ofIntermediate A (ethyl 15-chloro-9-(methoxymethyl)-2,4,8,10,11-pentaazatetracyclo[11.4.0.0²,⁶.0⁸,¹²]-heptadeca-1 (17),3,5,9,11,13,15-heptaene-5-carboxylate) was 72.58%. m/z calculated forC₁₇H₁₆ClN₅O₃[M+H]⁺: 374; Obtained: 374.1.

Synthesis of Intermediate B(15-chloro-9-(methoxymethyl)-2,4,8,10,11-pentaazatetracyclo[11.4.0.0²,⁶.0⁸,¹²]heptadeca-1(17),3,5,9,11,13,15-heptaene-5-carboxylic acid)

Intermediate A (0.4 gm, 1.07 mmol) was dissolved in mixture ofTHF/H₂O/MeOH (3.2/4.8/8.0 v/v mL). 0.05 gm, 2.14 mmol of LiOH was addedand the reaction mixture was stirred at room temperature for 3 h. Thereaction mixture was acidified with aq. 2N HCl solution, ppts werecollected and washed with DI water. After drying 0.36 gm of IntermediateB(15-chloro-9-(methoxymethyl)-2,4,8,10,11-pentaazatetracyclo[11.4.0.0²,⁶.0⁸,¹²]heptadeca-1(17), 3,5,9,11,13,15-heptaene-5-carboxylic acid) was obtained as a whitesolid. m/z calculated for C₁₅H₁₂ClN₅O₃ [M+H]⁺: 346; Obtained: 345.9.

Scheme 29 illustrates some selected examples using Intermediate A togenerate new analogs.

Synthesis of Compound 233

Acetoxime (0.22 gm, 0.31 mmol) was dissolved in anhydrous THF (0.5 mL).0.38 mL, 0.62 mmol of 1.6 M n-BuLi was added dropwise and reactionmixture stirred at 0-5° C. for 1 h in separate flask. A solution ofIntermediate A (0.05 gm, 0.13 mmol) in 1.0 mL of THF was added bycannula at 0-5° C. and the rxn was stirred for 16 h by gradually warmingat room temperature. LCMS indicated starting material and intermediatem/z: 374.1 (˜45/14%, two peak merged), m/z 401 (˜10%), m/z 402 (18%).

The reaction mixture was quenched with 0.03 mL of Conc. H₂SO₄, followedby 0.03 mL of DI water and refluxed for 2 h. LCMS indicated startingmaterial, product and intermediate m/z: 374.1 (˜43%), m/z 383 (˜40%),m/z 402 (17%).

The reaction mixture was concentrated under reduced pressure andneutralized with aq. NaHCO₃ solution, the ppts collected and washed withDI water. After drying gave 22.0 mg of crude ppts. Compound was purifiedby prep-TLC plate using 1:99 MeOH:CHCl₃.

Synthesis of Compound 238

Step: 1 Intermediate A (0.045 gm, 0.12 mmol) was dissolved in anhydroustoluene (3.0 mL). 0.05 mL, 0.25 mmol of aminoethanol (35.0 eq) was addedand reaction mixture was refluxed for 16 h. The toluene was evaporatedand reaction mixture was dissolved in DCM (25.0 mL). The DCM layer waswashed with brine followed by DI water, separated and dried overanhydrous MgSO₄. The evaporation of organic layer gave 38.3 mg of thecorresponding amide. LCMS indicated product formation m/z: 389

Step: 2 The above amide (0.038 gm, 0.09 mmol) was dissolved in dry DCM(2.0 mL). 0.026 mL, 0.2 mmol of DAST (2.0 eq) was added to the reactionmixture at 0° C. temperature and stirred for 1.5 h at 0° C. 0.065 gmsolid K₂CO₃ (4.8 eq) was added at 0° C. and reaction mixture was stirredfor 30 min. The reaction mixture was diluted with aq. NaHCO₃ solutionand extracted with DCM (15.0 mL×3). The organic layer was washed withbrine, separated and dried over anhydrous MgSO₄. The evaporation ofsolvent gave 36 mg of white solid product. m/z calculated forC₁₇H₁₅ClN₆O₂[M+H]⁺: 371; Obtained: 371.

Synthesis of Compound 239

Step: 1 Intermediate A (0.05 gm, 0.13 mmol) was dissolved in anhydroustoluene (3.0 mL). 0.28 gm, 2.67 mmol of aminoethanol (20.0 eq) was addedand reaction mixture was refluxed for 16 h. The toluene was evaporatedand reaction mixture was dissolved in DCM (25.0 mL). The DCM layer waswashed with brine followed by DI water, separated and dried overanhydrous MgSO₄. The evaporation of organic layer gave the amide. LCMSindicated product formation m/z: 431

Step: 2 The above amide (0.057 gm, 0.13 mmol) of was dissolved in dryDCM (2.0 mL). 0.035 mL, 0.3 mmol of DAST (2.0 eq) was added to thereaction mixture at 0° C. temperature and stirred for 1.5 h at 0° C.LCMS indicated product formation m/z 413. 0.088 gm solid K₂CO₃ (4.8 eq)was added at 0° C. and reaction mixture was stirred for 30 min. Thereaction mixture was diluted with aq. NaHCO₃ solution and extracted withDCM (15.0 mL×3). The organic layer was washed with brine, separated anddried over anhydrous MgSO₄. Concentration of the organic layer affordedproduct which was triturated with 20/80 Hex/EtOAc to give a solid whichwas collected by filtration and dried: 49.4 mg (89%).

Synthesis of Compound 243

Step: 1 Intermediate A (0.05 gm, 0.13 mmol) was dissolved in anhydroustoluene (3.0 mL). 0.02 mL, 2.67 mmol of the amino alcohol (20.0 eq) wasadded and reaction mixture was refluxed for 16 h. LCMS indicatedstarting material left. Xylene was placed (3.0 mL) and 10.0 eq of3-aminobutan-1-ol added and reaction mixture refluxed for 16 h. Finallytotal 40.0 eq of amino ethanol was required to convert all startingmaterial into product in refluxing xylene. The rxn mixture cooled to 0°C. and ppts filtered. The filtrate was extracted with DCM (15.0 mL×4).The DCM layer was washed with brine followed by DI water, separated anddried over anhydrous MgSO₄. The evaporation of organic layer gave thecorresponding amide. LCMS indicated product formation m/z: 403.

Step: 2 The above amide (0.054 gm, 0.13 mmol) was dissolved in dry DCM(2.0 mL). 0.05 mL, 0.33 mmol of DAST was added to the reaction mixtureat 0° C. temperature and stirred for 1.5 h at 0° C. LCMS indictedproduct formation. 0.09 gm solid K₂CO₃ was added at 0° C. and reactionmixture was gradually warmed to room temperature. The reaction mixturewas diluted with aq. NaHCO₃ solution and extracted with DCM (15.0 mL×3).The organic layer was washed with brine, separated and dried overanhydrous MgSO₄. The evaporation of solvent gave crude product.Purification was performed by prep TLC, Mobile Phase: 95:05, DCM:MeOH.m/z calculated for C₁₈H₁₇ClN₆O₂[M+H]⁺: 385; Obtained: 385.

Synthesis of Compound 244

Compound 243 from above (0.011 gm, 0.03 mmol) was dissolved in toluene(2.0 mL). 0.010 gm, 0.04 mmol of DDQ was added and reaction mixture wasstirred at 50° C. for 1 h. LCMS indicated starting material m/z 385 andlittle amount of product m/z 383. The rxn mixture was stirred at 60° C.for 3 h. LCMS indicated starting material m/z 385, product m/z 383. Therxn mixture was stirred at 70° C. for 2 h. LCMS indicated startingmaterial m/z 385, product m/z 383 and side product m/z 421. The reactionmixture was stirred at 40° C. for 16 h. LCMS indicated major amount ofproduct m/z 383 and little amount of side product m/z 421 and startingmaterial. The toluene was evaporated and crude product was purified byprep-TLC plate. Mobile phase DCM:MeOH, 95:05 v/v to obtain 4.4 mg ofproduct. m/z calculated for C₁₈H₁₅ClN₆O₂[M+H]⁺: 383; Obtained: 383.

Synthesis of Compound 249:

Compound 238 from above (0.016 gm, 0.05 mmol) was dissolved in toluene(2.0 mL). 0.015 gm, 0.07 mmol of DDQ was added and reaction mixture wasstirred at 50° C. for 1 h. LCMS indicated starting material m/z 371. Therxn mixture was stirred at 60° C. for 5 h. LCMS indicated startingmaterial m/z 371, product m/z 369 and undesired m/z 407. The rxn mixturewas stirred at 30° C. for 16 h. LCMS indicated starting material m/z371, product m/z 369 and side product m/z 407. The reaction mixture wasstirred at 65° C. for 4 h. LCMS indicated product m/z 369, side productm/z 407 and little amount of starting material. The toluene wasevaporated and crude product was purified by prep-TLC plate. Mobilephase DCM:MeOH, 95:05 v/v to obtain 2.3 mg of product. m/z calculatedfor C₁₇H₁₃ClN₆O₂[M+H]⁺: 369; Obtained: 369.

Synthesis of Compound 256

Step 1: Intermediate A (0.1 gm, 0.27 mmol) was dissolved in anhydrousTHE (3.0 mL). 0.67 mL, 0.67 mmol of 1.0 M solution of DIBAL in THE wasadded dropwise and reaction mixture stirred at 0-5° C. for 2 h. LCMSshows alcohol reduction product formation m/z 332. The reaction wasquenched with MeOH (1.0 mL), followed by water (0.5 mL). The saturatedsolution of NaHCO₃ was added and ppts were filtered through celite bed.The product was extracted using DCM (25.0 mL×3). The combined DCM layerswas washed with brine, separated and dried over anhydrous Na₂SO₄. Theevaporation of solvent gave 46.1 mg of[15-chloro-9-(methoxymethyl)-2,4,8,10,11-pentaazatetracyclo[11.4.0.0²,⁶.0⁸,¹²]heptadeca-1(17), 3,5,9,11,13,15-heptaen-5-yl]methanol as a solid product, Yield51.9%. m/z calculated for C₁₅H₁₄ClN₅O₂[M+H]⁺: 332; Obtained: 332.

Step 2: The above alcohol (0.05 gm, 0.14 mmol) of was dissolved inanhydrous DCM (3.0 mL). 0.09 gm of Dess-Martin Periodinane was added andreaction mixture was stirred at room temperature for 2 h. LCMS showsproduct formation m/z 330. The reaction was quenched with 1N NaOHsolution (2-mL). The saturated solution of NaHCO₃ was added and theproduct was extracted using DCM (20.0 mL×3). The combined DCM layers waswashed with brine, separated and dried over anhydrous Na₂SO₄. Theevaporation of solvent gave desired aldehyde(15-chloro-9-(methoxymethyl)-2,4,8,10,11-pentaazatetracyclo[11.4.0.0²,⁶.0⁸,¹²]heptadeca-1(17), 3,5,9,11,13,15-heptaene-5-carbaldehyde) as a solid product, YieldQuantitative. m/z calculated for C₁₅H₁₂ClN₅O₂ [M+H]⁺: 330; Obtained:330.

Step 3: 1.6 M n-BuLi solution in hexane (0.68 mL, 1.08 mmol) was addeddropwise into 1.4 mL, 0.86 mmol of trimethylsilyldiazomethane solutionin hexane dissolved in 3.0 mL of THE at −78° C. temperature. Thereaction mixture was stirred at −78° C. temperature for 30.0 min. Thealdehyde obtained in Step 2 (0.142 gm, 0.43 mmol) in solution in 3.0 mLof THE was added dropwise into the reaction mixture at −78° C.temperature and gradually warmed to room temperature. LCMS shows productformation m/z 326 and starting material m/z 330. The reaction mixturewas quenched with saturated NH₄Cl solution. The product was extractedusing DCM (15.0 mL×3). The combined DCM layers was washed with brine,separated and dried over anhydrous Na₂SO₄. The purification of crudeproduct was performed by ISCO Combiflash purification system, MobilePhase: Ethyl acetate/Hexane. 19.0 mg of Compound 256 was obtained and71.6 mg of starting material was isolated. m/z calculated forC₁₆H₁₂ClN₅O [M+H]⁺: 326; Obtained: 326.

Synthesis of Compound 285

Compound 256 (0.025 gm, 0.08 mmol) was dissolved in degassed DMF (2.0mL). 0.03 mL, 0.23 mmol of iodobenzene was added to the reaction mixturefollowed by 0.06 mL, 0.41 mmol of TEA. The reaction mixture was stirredat room temperature. 0.04 gm, 0.04 mmol of Pd(PPh₃)₄ and 0.003 gm, 0.015mmol of CuI mixture was added to the reaction mixture and stirred for 16h. LCMS shows product formation m/z 402. The reaction mixture wasdiluted with DI water. The product was extracted using DCM (10.0 mL×3).The combined DCM layers was washed with brine, separated and dried overanhydrous Na₂SO₄. The crude reaction mixture was purified throughprep-TLC plate. Mobile Phase: EtOAc/MeOH. m/z calculated for C₂₂H₁₆ClN₅O[M+H]⁺: 402; Obtained: 402.

Synthesis of Compound 254

Isobutyronitrile (10.0 gm, 144.70 mmol) was dissolved in EtOH:Water(150:50 mL, v/v), followed by addition of 10.0 gm, 144.70 mmol ofhydroxylamine hydrochloride and 20.0 gm, 144.70 mmol of K₂CO₃. Thereaction mixture was refluxed at 80° C. for 6 h. The solvent wasevaporated under reduced pressure and the resulting solid was treatedwith 150 mL of ethanol, sonicated, filtered and washed with 100 mL ofethanol. The combined filtrate was evaporated under reduced pressure andazeotrope with toluene (25.0 mL×3) to afford 8.1 gm ofN′-hydroxy-2-methylpropimidamide as a colorless liquid slurry (54.8%yield). The above amide-oxime (1.37 gm, 13.38 mmol) was azeotroped withtoluene (10 mL×5) before use and dissolved in 20.0 mL of anhydrous THF.0.27 gm, 6.69 mmol of NaH was added in three portion to the reactionmixture at 0° C. and stirred at ambient temperature for 30.0 min. 0.5gm, 1.34 mmol of Intermediate A was added and reaction mixture wasstirred for 45.0 min at ambient temperature and refluxed at 67° C. for90.0 min. The solvent was evaporated under reduced pressure andresulting yellow paste treated with 25.0 mL of aq. saturated NaHCO₃solution. The ppts were filtered through funnel and washed with water10.0 mL and hexane 10.0 mL to afford 0.380 gm solid (69.1% yield). m/zcalculated for C₁₉H₁₈ClN₇O₂[M+H]⁺: 412.0; Obtained: 412.1.

Synthesis of Compound 215

The alcohol [15-chloro-9-(methoxymethyl)-2,4,8,10,11-pentaazatetracyclo[11.4.0.0²,⁶.0⁸,¹²] heptadeca-1 (17),3,5,9,11,13,15-heptaen-5-yl]methanol (prepared in Compound 256, Step 1)(34 mg, 0.1025 mmol) was suspended in dry THF (2 mL). HMPA (36.7 mg,0.205 mmol) was added followed by ethyl iodide (0.33 mL) and NaH (41 mgof 60% suspension in oil). The reaction was stirred at RT for 5 min,then heated to 70° C. overnight. The mixture was cooled and partitionedbetween EtOAc and brine. The organic phase was dried and concentrated toafford an oil which was purified by column chromatography (0% to 10%MeOH in DCM) to give 3.7 mg of compound 215 as an oil.

Synthesis of Compound 274

[15-chloro-9-(methoxymethyl)-2,4,8,10,11pentaazatetracyclo[11.4.0.0²,⁶.0⁸,¹²]heptadeca-1 (17),3,5,9,11,13,15-heptaen-5-yl]methanol (0.02 gm, 0.06 mmol) was dissolvedin anhydrous THE (3.0 mL). 0.003 gm of NaH was added and reactionmixture was stirred at room temperature for 30.0 min. 0.012 mL, 0.12mmol of 2-bromopyridine was added dropwise and reaction mixture stirredat room temperature for 16 h. The reaction mixture was refluxed foradditional 2 h. LCMS shows m/z 409. The reaction was concentrated underreduced pressure and diluted with saturated solution of NaHCO₃. Theproduct was extracted using DCM (10.0 mL×4). The combined DCM layers waswashed with brine, separated and dried over anhydrous Na₂SO₄.Purification was performed by prep TLC, Mobile Phase: 95:05, DCM:MeOH.˜1.0 mg of product obtained. m/z calculated for C₂₀H₁₇ClN₆O₂[M+H]⁺: 409;Obtained: 409.

Scheme 30 illustrates some selected examples using Intermediate B togenerate new analogs.

Synthesis of Compound 234

Intermediate B (0.043 gm, 0.12 mmol), 0.3 mmol of EDC.HCl and 0.048 gm,0.31 mmol of HOBt hydrate were dissolved in THF/DCM (1:1, v/v 1.5 mL),followed by addition of 0.09 mL, 0.62 mmol of trimethylaniline and 0.016mL, 0.25 mmol of propargylamine. The reaction mixture was stirred atroom temperature for 16 h. The reaction mixture was diluted with aq.Ammonium chloride and extracted with ethylacetate. Combined layers werewashed with brine, separated and dried over anhydrous MgSO₄. Evaporationof organic layer gave crude product ˜13.0 mg. The crude product waspurified through preparative TLC plate, Mobile Phase: 5:95, MeOH,Ethylacetate. m/z calculated for C₁₈H₁₅ClN₆O₃[M+H]⁺: 383; Obtained:383.1

Synthesis of Compound 240

Step 1 Intermediate B (0.05 gm, 0.15 mmol) was dissolved in dry DCM (2.0mL). 0.05 mL, 0.36 mmol of trimethylamine (2.5 eq), followed by 0.024mL, 0.29 mmol of oxalylchloride (2.0 eq) were added and reaction mixturestirred for 60 min at room temperature. 0.076 mL, 0.7 mmol ofamino-alcohol (5.0 eq) was added to reaction mixture at 0° C. andstirred for 2.5 h. The reaction mixture was diluted with aq. solution ofNaHCO₃ and extracted with DCM (15.0 mL×3). The combined organic layerswere washed with brine, separated and dried over anhydrous MgSO₄. Theevaporation of organic layer gave 54.1 mg the amide. LCMS indicatedproduct formation m/z: 431

Step 2 (2S)-2-amino-3-methylbutyl15-chloro-9-(methoxymethyl)-2,4,8,10,11-pentaazatetracyclo[11.4.0.0²,⁶.0⁸,¹²]heptadeca-1(17), 3,5,9,11,13,15-heptaene-5-carboxylate (0.027 gm, 0.06 mmol) wasdissolved in dry DCM (2.0 mL). 0.016 mL, 0.13 mmol of DAST was added tothe reaction mixture at 0° C. temperature and stirred for 3 h at 0-5° C.LCMS indicted product formation. 0.04 gm solid K₂CO₃ was added at 0° C.and reaction mixture was gradually warmed to room temperature. Thereaction mixture was diluted with aq. NaHCO₃ solution and extracted withDCM (15.0 mL×3). The organic layer was washed with brine, separated anddried over anhydrous MgSO₄. The evaporation of solvent gave crudeproduct. Purification was performed by prep TLC, Mobile Phase: 95:05,DCM:MeOH. 23.7 mg of solid product was obtained. Mass. m/z calculatedfor C₂₀H₂₁ClN₆O₂[M+H]⁺: 413; Obtained: 413.

Synthesis of Compound 246

Compound 240 was converted to Compound 246 using DDQ, Toluene at 50 C inan analogous manner to Compound 245 to give 5.5 mg (37%) of Compound246. LCMS indicated product formation m/z: 411.

Synthesis of Compound 242

Step 1: Intermediate B (0.025 gm, 0.07 mmol) was dissolved in dry DCM(2.0 mL). 0.03 mL, 0.21 mmol of trimethylamine (3.0 eq), followed by0.015 mL, 0.18 mmol of oxalylchloride (2.5 eq) were added and reactionmixture stirred for 60 min at room temperature. 0.05 gm, 0.36 mmol of(R,S)-2-amino-2-phenylethan-1-ol (5.0 eq) was added to reaction mixtureat 0° C. and stirred for 2.5 h at room temperature. The reaction mixturewas diluted with aq. solution of NaHCO₃ and extracted with DCM (15.0mL×3). The combined organic layers were washed with brine, separated anddried over anhydrous MgSO₄. The evaporation of organic layer gave thedesired amide. LCMS indicated product formation m/z: 465

Step 2: The above amide (0.034 gm, 0.07 mmol) was dissolved in dry DCM(2.0 mL). 0.03 mL, 0.22 mmol of DAST was added to the reaction mixtureat 0° C. temperature and stirred at 0° C. for 1.5 h. LCMS indicatedproduct formation. 0.05 gm solid K₂CO₃ was added at 0° C. and reactionmixture was gradually warmed to room temperature. The reaction mixturewas diluted with aq. NaHCO₃ solution and extracted with DCM (15.0 mL×3).The organic layer was washed with brine, separated and dried overanhydrous MgSO₄. The evaporation of solvent gave crude product.Purification was performed by prep TLC, Mobile Phase: 95:05, DCM:MeOH.m/z calculated for C₂₃H₁₉ClN₆O₂[M+H]⁺: 447; Obtained: 447.

Synthesis of Compound 245

Compound 242 (0.015 gm, 0.03 mmol) was dissolved in toluene (1.5 mL).0.009 gm, 0.04 mmol of DDQ was added and reaction mixture was stirred at50° C. for 1.5 h. LCMS indicated starting material m/z 447 and productm/z 445 in 1:3 ratio. 0.005 gm, 0.022 mmol of DDQ was further added andrxn mixture was stirred at 50° C. for 1.5 h. starting material m/z 447and product m/z 445 in 1:6 ratio. The reaction mixture was stirred atroom temperature for 16 h. Purification was performed by prep TLC,Mobile Phase: 95:05, DCM:MeOH. The band with m/z: 445 was isolated and9.3 mg of solid compound was obtained (Yield 62.4%). m/z calculated forC₂₃H₁₇ClN₆O₂[M+H]⁺: 445; Obtained: 445.

Synthesis of Compound 237

Step 1: Intermediate B (0.025 gm, 0.07 mmol) was dissolved in dry DCM.0.009 mL, 0.02 mL, 0.14 mmol of trimethylamine, followed by 0.11 mmol ofoxalylchloride were added and reaction mixture stirred for 30 min atroom temperature. 0.028 mL, 0.36 mmol of 3-amino-1-propanol was added toreaction mixture at 0° C. and stirred for 2.5 h and then concentrated.LCMS indicated product formation m/z: 403, little starting materialleft.

Step 2: The crude amide from Step 1 (0.018 gm, 0.045 mmol) was dissolvedin dry DCM (2.0 mL). 0.012 mL, 0.09 mmol of DAST was added to thereaction mixture at −78° C. temperature and gradually warmed to 0° C.0.03 gm solid K₂CO₃ was added at −78° C. and reaction mixture wasgradually warmed to room temperature. The reaction mixture was dilutedwith aq. NaHCO₃ solution and extracted with DCM (15.0 mL×3). The organiclayer was washed with brine, separated and dried over anhydrous MgSO₄.The evaporation of solvent gave 14.7 mg of compound 237 as a white solidproduct. m/z calculated for C₁₈H₁₇ClN₆O₂[M+H]⁺: 385; Obtained: 385.1.

Synthesis of Compound 263

Step 1: Intermediate B (0.03 gm, 0.09 mmol), 0.034 gm, 0.17 mmol ofEDC.HCl and 0.027 gm, 0.17 mmol of HOBt.xH₂O were dissolved in anhydrousDCM (2.5 mL). 0.024 gm, 0.17 mmol of R-(−)-2-Phenylglycinol was addedand reaction mixture was stirred for 6 h at room temperature. LCMSindicated product formation m/z 464.9. The rxn mixture was diluted withDI water and extracted with DCM (10.0 mL×3). The combined DCM layerswere washed with brine, separated and dried over anhydrous Na₂SO₄. Theevaporation of organic layer gave crude product. A liquid syrup wasobtained. m/z calculated for C₂₃H₂₁ClN₆O₃ [M+H]⁺: 465; Obtained: 464.9.

Step 2: The above amide (0.04 gm, 0.086 mmol) of was dissolved in dryDCM (2.0 mL). 0.03 mL, 0.21 mmol of DAST was added and reaction mixturewas stirred at 0° C. temperature for 2 h. LCMS indicated productformation m/z 446.9. 0.06 gm solid K₂CO₃ was added at 0° C. and reactionmixture was gradually warmed to room temperature. The reaction mixturewas diluted with aq. NaHCO₃ solution and extracted with DCM (15.0 mL×3).The organic layer was washed with brine, separated and dried overanhydrous Na₂SO₄. The evaporation of solvent gave crude product.Purification was performed by prep TLC, Mobile Phase: 95:05, DCM:MeOH.25.0 mg of solid product was obtained. m/z calculated forC₂₃H₁₉ClN₆O₂[M+H]⁺: 447; Obtained: 446.9.

Synthesis of Compound 264

Step 1: Intermediate B (0.03 gm, 0.09 mmol), 0.034 gm, 0.17 mmol ofEDC.HCl and 0.027 gm, 0.17 mmol of HOBt.xH₂O were dissolved in anhydrousDCM (2.5 mL). 0.024 gm, 0.17 mmol of S-(+)-2-Phenylglycinol was addedand reaction mixture was stirred for 6 h at room temperature. LCMSindicated product formation m/z 464.9. The rxn mixture was diluted withDI water and extracted with DCM (10.0 mL×3). The combined DCM layerswere washed with brine, separated and dried over anhydrous Na₂SO₄. Theevaporation of organic layer gave crude product. A liquid syrup wasobtained. m/z calculated for C₂₃H₂₁ClN₆O₃[M+H]⁺: 465; Obtained: 464.9.

Step: 2: The above amide (0.04 gm, 0.086 mmol) was dissolved in dry DCM(2.0 mL). 0.03 mL, 0.21 mmol of DAST was added and reaction mixture wasstirred at 0° C. temperature for 2 h. LCMS indicated product formationm/z 446.9. 0.06 gm solid K₂CO₃ was added at 0° C. and reaction mixturewas gradually warmed to room temperature. The reaction mixture wasdiluted with aq. NaHCO₃ solution and extracted with DCM (15.0 mL×3). Theorganic layer was washed with brine, separated and dried over anhydrousNa₂SO₄. The evaporation of solvent gave crude product. Purification wasperformed by prep TLC, Mobile Phase: 95:05, DCM:MeOH. 26.4 mg of solidproduct was obtained. m/z calculated for C₂₃H₁₉ClN₆O₂[M+H]⁺: 447;Obtained: 446.9.

Compounds 180, 181, and 182 were prepared using a synthetic procedurethat is similar to the one used for the synthesis of Compound 168 asdepicted in Scheme 27.

Compounds 183-193 were prepared using a synthetic procedure that issimilar to the one used for the syntheses of Compounds 169-179 asdepicted in Scheme 26.

Compounds 194 and 195 were prepared using a synthetic procedure that issimilar to the one depicted in Schemes 21 and 22.

Compounds 196-198, and 206 were prepared using a synthetic procedurethat is similar to the one depicted in Scheme 18a.

Compound 202 was prepared using a synthetic procedure that is similar tothe one used for the synthesis of Compound 129 as depicted in Scheme18a.

Compounds 199, 200, 204, and 205 were prepared using a syntheticprocedure that is similar to the one depicted in Scheme 18b.

Compounds 201 and 203 were prepared using a synthetic procedure that issimilar to the one depicted in Scheme 24.

Compounds 207-210 were prepared using a synthetic procedure that issimilar to the one depicted in Scheme 17.

The nitrile substituents in Compounds 207-210 were generated analogouslyto those transformations shown in Scheme 22.

Compounds 211-214 were prepared using a synthetic procedure that issimilar to the one depicted in Scheme 20.

Compound 255 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound254.

Compound 259 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound243.

Compound 260 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 30; similar to compound242.

Compound 261 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 30; similar to compound256.

Compound 265 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 30; similar to compound264.

Compound 266 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 30; similar to compound264.

Compound 267 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 30; similar to compound264.

Compound 268 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 30; similar to compound263.

Compound 270 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 30; similar to compound264.

Compound 271 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 30; similar to compound264.

Compound 275 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 30; similar to compound264.

Compound 276 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 30; similar to compound245.

Compound 278 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound233.

Compound 281 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound233.

Compounds 282, 283, 286, 287 were prepared from the appropriate startingmaterials using the synthetic routes described in Schemes 28 and 29;similar to compound 243.

Compound 288 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound256.

Compound 293 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound285.

Compounds 294, 295, and 296 were prepared from the appropriate startingmaterials using the synthetic routes described in Schemes 28 and 29;similar to compounds 243 and 244.

Compound 303 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound233.

Compound 304 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound264.

Compound 297 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound243.

Compound 307 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound285.

Compound 308 was prepared from the appropriate starting materials usingthe synthetic routes described in Scheme 28; similar to Intermediate A.

Compound 309 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound238.

Compound 310 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound285.

Compound 311 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound285.

Compound 312 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound244.

Compound 313 was prepared from the appropriate starting materials usingthe synthetic routes described in Schemes 28 and 29; similar to compound244.

Synthesis of Compounds 305 and 306

Compound 288 (0.015 gm, 0.042 mmol) was dissolved in anhydrous THE (3.0mL). 0.003 mL, 0.05 mmol of methyl iodide was added at −78° C.temperature, followed by 0.05 mL, 0.05 mmol of 1.0 M LDA solution. Thereaction mixture was stirred at −78° C. and gradually warmed at roomtemperature. LCMS shows product formation m/z 368 major, unreactedstarting material m/z 354 and dimethylated unknown product m/z 382.1.The reaction mixture was quenched with saturated NH₄Cl solution andextracted with EtOAC. Organic layer was dried and concentrated. Thepurification of crude reaction mixture was performed by prep-TLC plate,Mobile Phase: EtOAc:Hexane 75:25 v/v mL to isolate three bands. It wasfound through MS that 1^(st) band confirmed m/z 354 of startingmaterial, 2^(nd) band confirmed m/z 368 of mono methyl substitutedproduct Compound 305 and 3^(rd) band confirmed m/z 382.1 of dimethylsubstituted product Compound 306. ¹H NMR (CDCl₃) data confirmed the monomethyl substitution on Imidazole ring. Note: ¹H NMR data confirmedproducts formation and pure products isolation.

Compound 216 was prepared similarly as compound 129 in Scheme 18a. MS:[M+1]=395.

Compound 217 was prepared similarly as compound 129 in Scheme 18a. MS:[M+1]=381.

Synthesis of Compound 218

To 5-(ethoxycarbonyl)-16-methoxy-2,3,4,10,12-pentaazatetracyclo[11.4.0.0^(2,6).0^(8,12)]heptadeca-1 (17),3,5,8,10,13,15-heptaene-9-carboxylic acid from Scheme 27 (0.609 g, 1.65mmol) stirring in DMF (10 ml) at 0° C. was added NaHCO₃ (0.749 g, 8.9mmol) and NBS (0.793 g, 4.45 mmol). The reaction was allowed to proceedto ambient temperature overnight. The reaction was then diluted withEtOAc, cooled to 0° C., and sat. sodium thiosulfate was added carefullyunder stirring. After foaming stopped, organic layer was separated,washed with sat. NaHCO₃, brine, and dried over MgSO₄. Filtration andsolvent removal gave the crude bromide which was purified by flashcolumn chromatography using a gradient elution of 0 to 80% EtOAc inhexanes. 424.2 mg (64%) was obtained as a yellowish solid. MS:[M+1]=405.

To the bromide (286.7 mg, 0.709 mmol) from above in a thick walled rbfwas added CuI (121.5 mg, 0.638 mmol), trimethylsilyl acetylene (1.04 g,10.7 mmol), triethyl amine (0.717 g, 7.09 mmol),dicyclohexyl(2′,6′-dimethoxybiphenyl-2-yl) phosphine (0.349 g, 0.851mmol) and 1,4-dioxane (2.5 ml; degassed). The reaction vessel wasflushed with nitrogen gas, and bis(triphenylphosphine) palladium(II)dichloride (298.2 mg, 0.425 mmol) was added. The reaction mixture wasstirred at rt for 30 min then heated at 100° C. under sealed tubeconditions for 16 hrs, diluted with EtOAc, and washed with sat. NaHCO₃,brine, and dried (MgSO₄). Silica gel column chromatography of thefiltered and concentrated reaction mixture using a gradient of 0 to 100%EtOAc in hexanes gave 157.9 mg (53%) of the desired trimethylsilylacetylene product as a brownish solid. MS: [M+1]=422.

The trimethylsilyl alkyne obtained above (128.7 mg, 0.305 mmol) wastreated with lithium hydroxide (36.6 mg, 1.53 mmol) in a solvent mixtureof THE (0.9 ml), water (0.75 ml) and MeOH (0.15 ml) at rt for two hrs.The mixture was then acidified to pH 3-4 with dil. Hydrochloric acid,and extracted with EtOAc (3×). The remaining precipitate in the aq.Layer was found to be product and was collected by filtration, and wascombined with the product isolated from the organic layer to give 95.6mg of the acid as a yellowish solid.

To the acid (95.6 mg, 0.298 mmol) in THE (1.3 ml) and dichloromethane(1.3 ml) was added N,O-dimethylhydroxylamine hydrochloride (232.4 mg,2.38 mmol), EDC hydrochloride (456.7 mg, 2.38 mmol), HOBt hydrate (91.2mg), and triethyl amine (0.833 ml, 5.93 mmol). After 16 hrs stirring,the reaction was diluted with EtOAc, and washed with sat. NH₄Cl. Aq.Layer was separated and extracted with EtOAc (3×), combined organiclayer was washed with sat. NaHCO₃, brine, and dried (MgSO₄). Filtrationfollowed by solvent removal gave 104.8 mg of the amide as a yellowishsolid.

To the Weinreb amide from above (20.1 mg, 0.0552 mmol) stirring in anh.THE (0.8 ml) cooled in an ice-salt bath was added 4-fluorophenylmagnesium bromide solution (1M THF; 0.828 ml) slowly. The reactionmixture was stirred to ambient temperature over 4 hrs, then quenchedwith sat. NH₄Cl, extracted with EtOAc (3×), washed with sat. NaHCO₃,brine, and dried (MgSO₄). Prep. TLC of the filtered concentrated mixtureusing 5% MeOH in DCM gave 2.0 mg of Compound 218 as an off-white solid.MS: [M+1]=400.

Compound 219 was prepared similarly as compound 218 as depicted inScheme 32. MS: [M+1]=416.

Synthesis of Compound 220

5-benzoyl-9-ethynyl-16-methoxy-2,3,4,10,12-pentaazatetracyclo[11.4.0.0^(2,6).0^(8,12)]heptadeca-1 (17), 3,5,8,10,13,15-heptaene (90.3mg, 0.237 mmol; obtained similarly as 218, was stirred in THF (1.5 ml)at rt. NaBH₄ (26.8 mg, 0.71 mmol) was added. After 1 hr, the reactionwas quenched with NH₄Cl for 5 min, and extracted with EtOAc. Organiclayer was separated and washed with brine and dried over MgSO₄.Filtration and solvent removal in vacuo gave a clear viscous oil, whichwas treated with triethylsilane (241.9 mg, 2.08 mmol) andtrifluoroacetic acid (0.32 ml) in DCM (1.5 ml) for 3 hrs. The reactionmixture was placed on Rotovap for solvent removal, diluted with EtOAc,and washed with sat. NaHCO₃. Aq. Layer was separated and extracted withEtOAc, the combined organic layer was washed with brine, and dried overMgSO₄. Prep. TLC of the filtered concentrate using 2% MeOH in DCM/EtOAc(1:1) gave 2.5 mg of Compound 220 as a clear filmy solid. MS: [M+1]=370.

Compound 221 was prepared similarly as compound 220 as depicted inScheme 32. MS: [M+1]=384.

Synthesis of Compound 222

The cyano ester (407.1 mg, 1.16 mmol) was treated with lithium hydroxide(83.5 mg, 3.49 mmol) in a solvent mixture of THF (6 ml), water (5 ml)and MeOH (1 ml) at rt for 16 hrs, then concentrated in vacuo, acidifiedto pH 3-4 with dil. HCl, and cooled at 0° C. Precipitate was collectedby filtration, washed with small amount of water, and dried to give271.9 mg (73%) acid as a greyish solid. This acid (271.9 mg) wassuspended and stirred in THE (2 ml) at 0° C., to which was added boranedimethylsulfide solution (2M THF; 8.4 ml) dropwise. The reaction wasallowed to proceed to ambient temperature overnight, cooled in an icebath, quenched with MeOH (10 ml) for two hrs, and concentrated in vacuo.The resulting solid residue was partitioned between DCM and sat. NaHCO₃and stirred for 20 min. Aq. Layer was separated and extracted with DCM(3×). Combined organic layer was washed with brine and dried over MgSO₄.Filtration and solvent removal gave 137.8 mg of the crude alcoholproduct as a yellowish waxy solid. The alcohol from above (137.8 mg) wastreated with phosphorus oxybromide (256.3 mg, 0.894 mmol) in 1,4-dioxane(5 ml) at 100° C. for 3 hrs. Upon cooling in an ice bath, the reactionmixture was treated with sat. NaHCO₃ (15 ml) and EtOAc (15 ml) understirring conditions for about 20 min. The basic aq. Layer was separatedand extracted with EtOAc (2×). Combined organic layer was washed withbrine and dried over MgSO₄. Filtration and solvent removal in vacuo gavethe crude primary bromide as a solid paste which was stored in cold andused without further purification when needed.

The crude bromide from above (27.0 mg, 0.0727 mmol) was treated with4-fluorophenol (65.2 mg, 0.585 mmol) and cesium carbonate (47.4 mg,0.145 mmol) at rt for 16 hrs. The reaction mixture was diluted withEtOAc, washed with brine, and dried over MgSO₄. Prep. TLC of thefiltered concentrate using 5% MeOH in DCM/EtOAc (1:1) gave 1.2 mg ofCompound 222 as a yellowish solid. MS: [M+1]=403.

Compound 223 was prepared similarly as compound 222 as depicted inScheme 33. MS: [M+1]=403.

Compound 224 was prepared similarly as compound 222 as depicted inScheme 33. MS: [M+1]=385.

Compound 225 was prepared similarly as compound 222 as depicted inScheme 33. MS: [M+1]=464.

Ethyl1-(5-chloro-2-nitrophenyl)-5-(2-ethoxy-2-oxoethyl)-1H-1,2,3-triazole-4-carboxylate(21.2 g; obtained similarly as 14 in Scheme 11) was treated with tin(II) chloride hydrate (60 g) in a mixture solvent of EtOAc/EtOH (1:2,300 ml) at 70° C. for 3 hrs. HCl (40 ml; 37%) was added and heatingcontinued for 3 days. More tin (II) chloride hydrate (25 g) and HCl (15ml) added and heating continued for 2 days. The reaction was cooled,concentrated under reduced pressure to a brownish oil, diluted withEtOAc (250 ml), and carefully basified to pH 8-9 with sodium carbonatesolution. The aq. Layer was separated and extracted with EtOAcrepeatedly. Combined organic layer was washed with brine and dried overMgSO₄. Filtration and solvent removal followed by recrystallization inMeOH gave 3.3 g (51%) of the cyclized mono-ester as a yellowish solid.MS: [M+1]=307.

Preparation of Tert-Butyl Isocyanoacetate:

To a suspension of tert-butyl glycinate hydrochloride (10.0 g, 60 mmol)in DCM (200 ml) was added EDC.HCl (14.9 g, 78 mmol) and triethylamine(12.5 mL, 89.8 mmol). The reaction mixture was cooled down to −50° C.,formic acid (3.4 mL, 89.8 mmol) in DCM (10 mL) was added slowly. Thereaction mixture was stirred at −50° C. for one hour then at 4° C. for 3h. Water (150 ml) was added. After 30 min stirring, aq. Layer wasseparated and extracted with DCM (3×). Combined organic layer was washedwith brine and dried over MgSO₄. Filtration and solvent removal underreduced pressure gave 10 g (100%) of the formyl amide as a clear viscousoil. H¹NMR (CDCl₃) δ 8.23 (1H, s), 6.17 (1H, br s), 3.98 (2H, d, J=5.5Hz), and 1.48 (9H, s).

To a solution of formyl amide (10.5 g, 66 mmol) in DCM (180 mL) wasadded triethylamine (36.8 mL, 264 mmol). The solution was cooled in asalt-ice bath, and POCl₃ (7.4 mL, 79.2 mmol) was added slowly. Thereaction was stirred in the cold bath for one hr. Then sodium carbonate(7.7 g, 72.6 mmol) in water (90 ml) was added to the cold reactionmixture. After 15 min, cold bath was removed and stirring continued atambient temperature for one hr. Aq. Layer was separated and extractedwith DCM (3×). Combined organic layer was washed with brine and driedover MgSO₄. Filtration and solvent removal under reduced pressure gave7.9 g (84%) tert-butyl isocyanoacetate as a dark brown liquid. H¹NMR(CDCl₃) δ 4.12 (2H, s), and 1.51 (9H, s).

A solution of tert-butyl isocyanoacetate (1.51 g, 10.7 mmol) in DMF (43ml) was cooled to −50° C. under nitrogen atmosphere. Potassiumt-butoxide (1.05 g, 9.4 mmol; finely pressed) was added. After one hrstirring at −50° C., the 1,2,4-triazole intermediate (2.32 g, 6.48 mmol;prepared similarly as compound 20 in Scheme 11) was added to theresulting reddish clear solution, and the reaction was stirred toambient temperature overnight. Sat. NaHCO₃ (15 ml) was added, and thereaction mixture was extracted with diethyl ether (5×), washed withbrine, and dried (MgSO₄). Silica gel chromatography of the filteredconcentrate using a gradient of 0 to 100% EtOAc in hexanes gave 2.5 g(89%) of the imidazole t-butyl ester product as a yellowish solid. MS:[M+1−tBu]=374.

The imidazole t-butyl ester from above (1.1 g, 2.56 mmol) was treatedwith trifluoroacetic acid (13 ml) in DCM (13 ml) for 3 hr or until allstarting t-butyl ester was hydrolyzed. The reaction was thenconcentrated under reduced pressure. Residual TFA was removed withrepeated addition and evaporation of toluene. The acid product wasobtained as a dark brown viscous oily material, and was used withoutfurther purification. MS: [M+1]=374.

Ethyl16-chloro-9-cyano-2,3,4,10,12-pentaazatetracyclo[11.4.0.0^(2,6).0^(8,12)]heptadeca-1(17), 3,5,8,10,13,15-heptaene-5-carboxylate (477 mg, 1.34 mmol);obtained similarly as ethyl9-cyano-16-methoxy-2,3,4,10,12-pentaazatetracyclo[11.4.0.0^(2,6).0^(8,12)]heptadeca-1(17), 3,5,8,10,13,15-heptaene-5-carboxylate in Scheme 27) was treatedwith lithium hydroxide (80.5 mg, 3.36 mmol) in a solvent mixture of THF(6 ml), water (5 ml) and MeOH (1 ml) at rt for 16 hrs. The reaction wasconcentrated under reduced pressure, acidified to pH 3-4 with dil. HCl,and cooled to 0° C. Precipitate was collected by filtration, washed withsmall amount of water, and further dried to give 396.2 mg crude triazolocarboxylic acid product, MS: [M+1]=327.

To a suspension of the crude acid from above (396.2 mg) in anhydrous THE(7 ml) at 0° C. was added borane dimethylsulfide complex (10.9 ml; 2MTHF) dropwise. The reaction was allowed to proceed to ambienttemperature overnight, and was cooled to 0° C., then slowly quenchedwith MeOH. After 30 min stirring, the reaction mixture was concentratedin vacuo. The resulting slurry was treated with MeOH which wassubsequently removed in vacuo. This process was repeated several times.The resulting residue was then treated with 5% MeOH in DCM, and washedwith sat. NaHCO₃. Aq. Layer was extracted with DCM (3×), combinedorganic layer was washed with brine and dried over MgSO₄. Filtration andsolvent removal gave a mixture of the crude alcohol product ([M+1]=313)and the corresponding primary amide due to hydrolysis of the cyano group([M+1]=331). 388.8 mg of this crude mixture was obtained and was usedwithout further purification.

The alcohol mixture (388.8 mg) from above was treated with phosphorusoxybromide (2.02 g) in 1,4-dioxane (10 ml) at 100° C. for 8 hrs. Thereaction was cooled to 0° C., and carefully quenched with sat. NaHCO₃(15 ml). After 20 min stirring, the reaction mixture was extracted withEtOAc (3×), washed with brine, and dried over MgSO₄. Filtration andsolvent removal under reduced pressure gave the crude bromide as aviscous paste, which was used for the next step without furtherpurification.

Compound 226 was prepared similarly as Compound 222 in Scheme 33 usingthe bromide prepared from above. MS: [M+1]=389.

Compound 227 was prepared in a similar fashion as Compound 226, depictedin Scheme 34. MS: [M+1]=407.

Compound 228 was prepared in a similar fashion as Compound 226, depictedin Scheme 34. MS: [M+1]=407.

Synthesis of Compound 229:

The benzyl analog 229, shown in Scheme 35, was prepared similarly as thebenzyl compound 220 in Scheme 32. MS: [M+1]=411.

Synthesis of Compound 230:

The ketone analog 230, shown in Scheme 35, was prepared similarly asketone 218 in Scheme 32. MS: [M+1]=474.

Synthesis of Compound 231:

The benzyl analog 231, shown in Scheme 35, was prepared similarly as thebenzyl compound 220 in Scheme 32. MS: [M+1]=460.

Compound 63 (0.805 g, 1.78 mmol; from Scheme 18a) was treated withlithium hydroxide (0.128 g, 5.34 mmol) in a solvent mixture of THF (6ml), water (5 ml) and MeOH (1 ml) at rt for 16 hrs. The reaction wasthen concentrated in vacuo, acidified to pH 3-4 with dil. HCl. Resultingprecipitate was collected by filtration, washed with water and dried togive 0.638 g acid as a yellow solid. MS: [M+1]=424.

The acid from above (0.638 g, 1.5 mmol) was treated with NBS (1.61 g, 9mmol) and NaHCO₃ (1.51 g, 18 mmol) at rt for 16 hrs. The reactionmixture was cooled to 0° C., sat. sodium thiosulfate (aq.) was carefullyand slowly added. This was extracted with EtOAc (2×), washed with sat.NaHCO₃, brine, and dried over MgSO₄. Silica gel chromatography of thefiltered concentrate with a gradient of 0 to 100% EtOAc in hexanes gave0.580 g (72%) of the di-bromo product as a yellowish solid. MS:[M+1]=538.

Compound 232 was prepared similarly as Compound 55 in Scheme 18a, usingthe bromide prepared above. MS: [M+1]=439.

Compound 235 was prepared similarly as Compound 55 in Scheme 18a, usingthe bromide prepared above. MS: [M+1]=440.

Compound 236 The alkyne moiety was prepared similarly as Compound 161 inScheme 21. MS: [M+1]=384.

Compound 241 The alkyne moiety was prepared similarly as Compound 161 in

Synthesis of Compound 247:

The bromide ester (13.9 mg, 0.0344 mmol) was treated with lithiumhydroxide (10 mg) in a solvent mixture of THF (0.3 ml), water (0.25 ml)and MeOH (0.05 ml) at rt for 16 hrs. The reaction was then concentratedin vacuo, acidified to pH 3-4 with dil. HCl and cooled to 0° C.Resulting precipitate was collected by filtration, washed with water anddried to give 9.5 mg (74%) acid as a light brown solid. MS: [M+1]=377.To the acid from above (5.1 mg, 0.0136 mmol) stirring in DCM (0.15 ml)was added oxalyl chloride (8.6 mg, 0.0678 mmol), and DMF (5 ul). After 2hrs stirring, solvent and excess reagent was removed in vacuo. Resultingresidue was re-suspended in DCM (0.15 ml), cooled in an ice-salt bath,and ethanolic methyl amine (100 ul; 33%) was added dropwise. After 20min stirring, the reaction mixture was applied to a prep. TLC plate andproduct was isolated using 5% MeOH in DCM as eluent. 4.3 mg (81%)Compound 247 was obtained as a white solid. MS: [M+1]=390.

Compound 248 was prepared similarly as Compound 247, as depicted in

To the acid (108.0 mg, 0.335 mmol) suspended in DCM (2 ml) at 0° C. wasadded oxalyl chloride (170.1 mg, 1.34 mmol) slowly, followed by DMF (20ul). After bubbling stopped, ice bath was removed and the reaction wasallowed to proceed at rt for 2 hrs. Solvent and excess reagent wasremoved in vacuo. Resulting light brown solid was cooled to 0° C. NaBH₄solution (2.2 ml; 1.5M in methoxyethoxy ethane) was added. After 30 min,the reaction was quenched with 1N HCl (0.2 ml), and stirring continueduntil bubbling stopped. EtOAc (10 ml) and sat. NaHCO₃ (10 ml) was addedand this was stirred overnight. Aq. Layer was separated and extractedwith EtOAc (3×); combined organic layer was washed with brine and driedover MgSO₄. Filtration and solvent removal gave 97.0 mg (94%) of thealcohol as a yellowish solid. MS: [M+1]=309.

The alcohol from above (97.0 mg, 0.315 mmol) was treated withDess-Martin Periodinane (266.9 mg, 0.629 mmol) in DCM (2 ml) for 1 hr.The reaction mixture was diluted with DCM, washed with sat. NaHCO₃. Aq.Layer was separated and extracted with DCM (3×), combined organic layerwashed with brine, and dried over MgSO₄. Filtration and solvent removalunder reduced pressure gave quantitative yield of the crude aldehyde asa brownish solid, which was used without further purification.

Compound 250 was prepared similarly as compound 48 in Scheme 16 usingthe aldehyde from above, as depicted in Scheme 33. MS: [M+1]=362

Compound 251 was prepared similarly as compound 250, as depicted inScheme 33. MS: [M+1]=376.

Compound 252 was prepared similarly as compound 250, as depicted inScheme 33. MS: [M+1]=364.

Compound 253 was prepared similarly as compound 250, as depicted inScheme 33. MS: [M+1]=452.

The acid (16 in Scheme 15, X═OMe; 258.1 mg, 0.941 mmol) was treated withacetic acid (2 ml) at 120° C. for 5 hr. Solvent was then removed invacuo. Solid residue was treated in water (7 ml) with sonication,filtered, washed with water, and dried to give 158.4 mg (73%)decarboxylated product as a brownish solid. MS: [M+1]=231.

Compound 257 was prepared in a similarly fashion as compound 167 inScheme 11. MS: [M+1]=364.

Compound 258 was prepared in a similarly fashion as compound 167 inScheme 11. MS: [M+1]=336.

Synthesis of Compound 262

Benzyl triphenyl phosphonium bromide (29.0 mg, 0.0669 mmol) was stirredin THF (0.5 ml) cooled in a salt-ice bath. Sodium hydride (4.12 mg,0.103 mmol; 60% oil suspension) was added. After 20 min stirring,aldehyde (15.8 mg, 0.0515 mmol) was added. The reaction was allowed toslowly warm to rt over four hrs, then quenched with sat. NH₄Cl,extracted with EtOAc (3×), washed with brine, and dried over MgSO₄.Compound 262 was isolated by repeated prep. TLCs using 2% MeOH in DCM.1.1 mg was isolated as a white solid. MS: [M+1]=381.

The starting ester (76.4 mg, 0.235 mmol) was treated withN-bromosuccinamide (83.6 mg, 0.470 mmol) in acetonitrile (2.3 ml) at rtfor three days. To the reaction mixture was added sat. sodiumthiosulfate. After 15 min stirring, aq. Layer was separated andextracted with EtOAc (2×). Combined organic layer was washed with brineand dried over MgSO4. The bromide product was isolated by prep. TLCusing hexanes:EtOAc=1:3 as the eluting solvent. 50.2 mg (52%) wasobtained as a light brown foamy solid. MS: [M+1]=405.

To the bromide from above (24.1 mg, 0.0596 mmol) under nitrogen atm. wasadded phenyl boronic acid (10.3 mg, 0.083 mmol),tetrakis(triphenylphosphine)palladium(0) (6.9 mg, 0.006 mmol),dimethoxyethane (0.69 mL; degassed), and aq. Na₂CO₃ solution (77 ul;2M). The reaction was heated at 100° C. for 5 hrs, cooled to rt, dilutedwith EtOAc, washed with sat. NaHCO₃, brine, and dried over MgSO₄. Prep.TLC with hexanes:EtOAc=1:3 gave 17.2 mg (72%) Suzuki coupling product asa yellowish amorphous material. MS: [M+1]=402.

Syntheses of Compound 272, 273 and 277:

Compound 272 was prepared similarly as compound 167 in Scheme 11,starting from the imidazole ester above. MS: [M+1]=440.

Compound 273 was prepared similarly as compound 167 in Scheme 11,starting from the imidazole ester above. MS: [M+1]=412.

Compound 277 was prepared similarly as compound 167 in Scheme 11. MS:[M+1]=378.

Compound 279 was prepared via Suzuki coupling in a similar fashion asdetailed above (see Scheme 37). MS: [M+1]=436.

Synthesis of Compound 280:

To compound 267 (11.7 mg, 0.0274 mmol) under nitrogen atmosphere wasadded dicyclohexyl[2-(2,4,6-triisopropylphenyl) phenyl]phosphane (7.8mg, 0.0164 mmol), cesium carbonate (22.3 mg, 0.0685 mmol), andacetonitrile (0.30 ml). The reaction flask was flushed with nitrogengas, and dichlorobis(acetonitrile)palladium (II) (1.42 mg, 0.0055 mol)was added. After stirring at rt for 30 min, trimethylsilyl acetylene(80.7 mg, 0.822 mmol) was added, and the reaction was heated at 90° C.for 5 hrs, cooled to rt, diluted with EtOAc, and washed with sat.NaHCO₃. Aq. Layer was separated and extracted with EtOAc (2×), combinedorganic layer was washed with brine and dried over MgSO₄. Prep. TLC ofthe filtered concentrate using 5% MeOH in DCM/EtOAc (1:1) gave 4.1 mgtrimethylsilyl acetylene derivative as a yellowish solid. MS: [M+1]=489.

The trimethylsilyl acetylene (4.1 mg, 0.0084 mmol) from above wastreated with potassium carbonate (1.2 mg, 0.0084 mmol) in methanol (0.2ml) at rt for 3 hrs. Prep. TLC using 7% MeOH in DCM/EtOAc (1:1) aseluting solvent gave 1.6 mg Compound 280 as a yellowish solid. MS:[M+1]=417.

Syntheses of Compound 284, 301 and 302:

Compound 284 was prepared similarly as compound 280, starting fromcompound 240. MS: [M+1]=403.

Compound 301 was prepared similarly as compound 280 starting fromcompound 264. MS: [M+1]=437.

Compound 302 was prepared similarly as compound 280 starting fromcompound 245. MS: [M+1]=435.

Syntheses of Compound 289, 290, 291 and 292:

Compound 289 was prepared similarly as compound 263 as depicted inScheme 30. MS: [M+1]=399.

Compound 290 was prepared similarly as compound 263 as depicted inScheme 30. MS: [M+1]=399.

Compound 291 was prepared similarly as compound 243 as depicted inScheme 29. MS: [M+1]=337.

Compound 292 was prepared similarly as compound 243 as depicted inScheme 29. MS: [M+1]=337.

Synthesis of Compound 298:

The ester (107.9 mg, 0.264 mmol) in THE (2.4 ml) was treated withlithium borohydride solution (0.264 ml; 2M THF) at 0° C. The reactionwas allowed to warm to ambient temperature over 4 hrs, then quenchedwith sat. NaHCO₃ slowly, extracted with EtOAc (4×), washed with brine,and dried over MgSO₄. Filtration and solvent removal gave 77.3 mg (86%)alcohol as a yellowish solid.

Alcohol from above (16.4 mg, 0.0448 mmol) was treated with phosphorusoxybromide (25.7 mg, 0.0895 mmol) in 1,4-dioxane (0.5 ml) at 95° C. for3 hrs. The reaction was then cooled to 0° C., quenched with sat. NaHCO₃(5 ml) for 20 min, and extracted with EtOAc (3×), washed with brine, anddried over MgSO₄. Filtration and drying gave 16.6 mg yellowish solidwhich was dissolved in anhydrous MeOH (18 ul) and THE (0.35 ml). Thiswas cooled to 0° C., and NaH (9.2 mg; 60% suspension) was added. After 2hrs stirring at 0° C., the reaction was quenched with sat. NaHCO₃,extracted with EtOAc (3×), washed with brine, and dried over MgSO₄.Prep. TLC using 10% MeOH in DCM gave 0.8 mg Compound 298 as a yellowishsolid. MS: [M+1]=381.

The starting alcohol (616 mg) was converted to the corresponding bromideas described earlier (see Scheme 21). The resulting crude bromide wasdissolved in anhydrous methanol (23 ml), and cooled to 0° C. NaH (932mg; 60% suspension) was added portionwise. After bubbling stopped, thereaction mixture was heated to reflux for 30 min, then cooled to rt, andtreated with 2N HCl (11 ml). Resulting precipitate was collected byfiltration, and the desired methyl ether was isolated by silica gelchromatography, using a gradient elution of 0 to 10% MeOH in DCM. 217 mgwas collected as a yellowish solid. MS: [M+1]=279.

Syntheses of Compounds 299 and 300

Compound 299 was prepared similarly as Compound 289, using the methylether intermediate above. MS: [M+1]=461.

Compound 300 was prepared similarly as Compound 289, using the methylether intermediate above. MS: [M+1]=385.

Compounds 180-313 were characterized by MS and ¹H NMR. The MScharacterization is summarized below in Table 5.

TABLE 5 MS characterization of Compounds 180-313, Observed MS Cmp No.Structure (M + 1) 180

460 181

460 182

442 183

502 184

502 185

459 186

396 187

410 188

476 189

476 190

486 191

403 192

441 193

453 194

440 195

458 196

403 197

389 198

384 199

426 200

414 201

450 202

443 203

485 204

436 205

388 206

412 207

369 208

403 209

403 210

370 211

347 212

423 213

441 214

437 215

360 216

395 217

381 218

400 219

416 220

370 221

384 222

403 223

403 224

385 225

464 226

389 227

407 228

407 229

411 230

474 231

460 232

439 233

383 234

383 235

440 236

384 237

385 238

371 239

413 240

413 241

385 242

447 243

385 244

383 245

445 246

411 247

390 248

430 249

369 250

362 251

376 252

364 253

452 254

426 255

398 256

326 257

364 258

336 259

399 260

461 261

340 262

381 263

447 264

447 265

461 266

467 267

427 268

475 270

475 271

461 272

440 273

412 274

409 275

461 276

473 277

378 278

397 279

436 280

417 281

411 282

385 283

385 284

403 285

402 286

413 287

413 288

354 289

399 290

399 291

337 292

337 293

416 294

385 295

385 296

383 297

399 298

381 299

461 300

385 301

437 302

435 303

451 304

515 305

368 306

382 307

434 308

442 309

439 310

444 311

430 312

451 313

513

Implementing reactions similar and analogous to those shown in Schemes 1through 37, the following compounds are also specifically contemplatedin this application

Example 105: Assessing α5-Containing GABA_(A) Receptor (GABAAR) PositiveAllosteric Modulator Activity

Step 1: Establish clones of GABAAR subunits (α5, β3, γ², α1, α2 and α3)and prepare the corresponding cRNAs: Human clones of GABA_(A)-R α5, β3,γ2, α1, α2 and α3 subunits are obtained from commercial resources (e.g.,OriGene, http://www.origene.com and Genescript,http://www.genescript.com). These clones are engineered into pRC, pCDM,pcDNA, and pBluescript KSM vector (for oocyte expression) or otherequivalent expression vectors. Conventional transfection agents (e.g.,FuGene, Lipofectamine 2000, or others) are used to transiently transfecthost cells.

Step 2—Functional GABAAR Assay of α5β3γ2, α1β3γ2, α2β3γ2, and α3β3γ2,subtypes in Xenopus oocyte expression system: cRNAs encoding α5, β3, γ2,α1, α2 and α3 subunits are transcribed in vitro using T3 mMESSAGEmMACHINE Kit (Ambion) and injected (in a ratio of α:β:γ=2:2:1 or otheroptimized conditions) into oocytes freshly prepared from Xenopus laevis.After two days of culturing, GABA-gated Cl− currents from oocytes areperformed using TEVC setups (Warner Instruments, Inc., Foster City,Calif.). GABA, benzodiazepine, and diazepam are used as referencecompounds to validate the system.

Step 3—Evaluate test compounds for positive allosteric modulatoractivity on the α5β3γ2 subtype and test off-target activity on the α1 toα3 coupled β3γ2 subtypes when the EC50=5 μM selectivity cut-off isreached: The GABA-gated Cl− current from oocytes are measured in theTEVC setup in the presence of the test compounds. The positiveallosteric modulator activity of each the test compounds is tested in a5-point dose-response assay. The test compounds include some referencecompounds (literature EC50 values for the α5β3γ2 subtype are in therange of 3-10 μM). EC50s in the α5β3γ2 subtype are obtained for eachcompound. If the EC50 in α5β3γ2 is <5 M, then the EC50 of the otherthree subtypes (α1β2γ2, α2β3γ2, and α3β3γ2) is further determinedindividually in order to test for selectivity of the compounds in theα5β3γ2 subtype over other subtypes.

Step 4 Evaluate further test compounds on the α5β3γ2 subtype and testoff-target activities when the EC50=0.5 μM selectivity cut-off isreached: The second batch of test compounds are tested using the samestrategy, but with a lower EC50 cutoff (0.5 μM). Again, the EC50s of theα5β3γ2 subtype for each of the compounds is determined. The α1 to α3coupled β3γ2 subtypes are tested only if the EC50 for the α5-containingreceptor is <0.5 μM.

Example 106: Evaluating Compounds for Binding and Positive AllostericModulator Activity on the GABA_(A) α5 Receptors (A) Binding Activity ofTest Compounds on GABA_(A)R

Tissue culture and Membrane Preparation: The binding was performed onLtk cells stably expressing GABA_(A) receptors: α1β1γ2, α2β3γ2, α3β3γ2and α5β3γ2 (provided by Merck Co., NJ, USA). Cells were seeded in 100 mmculture plates in DMEM/F12 medium containing 10% serum and antibioticsin 5% CO2 and allowed to grow for 1-2 days. GABAAR expression was theninduced by dexamethasone as follows: 0.5 μM for 1 day for α5 containingand 2 μM for 3 days for α1, α2 and α3 containing GABA_(A)Rs. Afterinduction, cells were collected by scraping into Dulbecco's Phosphatebuffered saline (DPBS, pH 7.4, Invitrogen, Carlsbad, Calif., USA) andcentrifuged at 150×g for 10 min. The pellet was washed twice byre-suspension and centrifugation. The cell pellets from at least 5different preps were combined, suspended in the binding assay buffer (50mM KH2PO4; 1 mM EDTA; 0.2 M KCl, pH 7.4) and membranes prepared bysonication (3-5 times, 30 sec) using Branson Sonifier 150 (G. Heinmann,Germany). Protein content was determined using BCA assay (Bio-Rad Labs,Reinach, Switzerland) with Bovine Serum Albumin (Sigma Aldrich, St.Louis, Mo., USA) as the standard. Aliquots were prepared and stored at−20° C. for further use in binding assays.

Ligand Binding: Saturation binding curves were obtained by incubatingmembranes with increasing concentrations (0.01-8 nM) of [3H]Rol5-1788(Flumazepil, 75-85 Ci/mmol, PerkinElmer, MA, USA), with nonspecificbinding measured in the presence of 10 μM diazepam. Inhibition of[3H]Rol5-1788 binding of the test compounds was performed atconcentrations of the radioligand at or lower than the K_(d) values forα1, α2, α3 and α5 containing GABA_(A)Rs determined from the saturationcurves.

All binding assays were performed for 1 h at 4° C. in assay buffer. Thetotal assay volume was 0.5 ml containing 0.2 mg/ml protein for α5 and0.4 mg/ml for α1, α2, and α3 containing GABA_(A)R membranes. Incubationswere terminated by filtration through GF/B filters using a 24-CellHarvestor (Brandel, Gaithersburg, Md., USA) followed by 3 washes withice-cold assay buffer. Filters were transferred to scintillation vials,5 ml scintillation liquid added, vortex-mixed and kept in dark. Nextday, radioactivity was obtained using a scintillation counter (BeckmanCoulter, Brea, Calif., USA). All assays were performed in triplicate.

Data Analyses: Saturation and inhibition curves were obtained usingGraphPad Prism software (GraphPad Software, Inc., CA, USA). Theequilibrium dissociation constants (K_(i) values) of the unlabeledligand were determined using Cheng-Prusoff equationK_(i)=IC₅₀/(1+S/K_(d)), where IC₅₀ is the concentration of unlabeledligand that inhibits 50% of [³H] ligand binding, S is the concentrationof radioligand and K_(d) is the equilibrium dissociation constant of theradioactive ligand. A log range of the compounds (1 nM-10 μM) was usedto determine the K_(i) values which are presented as Mean±SD fromtriplicate assays.

(B) Positive Allosteric Modulator Activity of Test Compounds on α5β2γ2Subtype GABA_(A)R

Compounds of the present invention were initially screened at 100 nM fortheir ability to potentiate an EC₂₀ concentration of GABA in oocytescontaining GABA_(A) receptors (α5β2γ2), using a protocol essentiallysimilar to the one presented above.

On day 1, 1 ng/32 nL of GABA_(A) α5β2γ2 cDNA was injected into oneoocyte. Test starts on day 2. The cDNA injected to the oocytes was a mixof alpha, beta and gamma, their ratio is 1:1:10 (by weight) and thetotal weight of the mixed 3 subunits to be injected in one oocyte was 1ng in 32 nl volume. The injected oocytes can also be tested on day 3. Insuch case, the cDNA amount injected to the oocytes should be reduced by20%.

Compounds of the present invention were tested using the followingprocedures.

GABA Dose-Response

1). 8 oocytes were placed in 8 chambers of OpusXpress and superfusedwith Modified Barth's Saline (MBS) at 3 mL/min. Glass electrodesback-filled with 3M KCl (0.5-3 megaohms) were used. Membrane potentialof oocytes was voltage-clamped at −60 mV.2). Average EC₂₀ GABA obtained from previous tests were applied forfive-six times to stabilize oocytes. Oocytes were washed with MBS for5-10 min between each GABA applications.3). Run GABA dose-response to obtain EC₂₀ GABA value.

Control Test (Diazepam or Methyl 3,5-Diphenylpyridazine-4-Carboxylate)

1). New oocytes was used to run new test.2). EC₂₀ GABA was applied for five-six times to stabilize oocytes.Oocytes were washed with MBS for 5-10 min between each GABAapplications.3). EC₂₀ GABA was applied to obtain current (I_(GABA)). Oocytes werewashed with MBS for 5-10 min.4). 1 μM diazepam or methyl 3,5-diphenylpyridazine-4-carboxylate waspre-applied for sec, followed by co-application of 1 μM diazepam ormethyl 3,5-diphenylpyridazine-4-carboxylate and EC₂₀ GABA to obtainI_(test). I_(test) was divided by I_(GABA) to obtain potentiation (%).

Test Compounds at Multiple Doses

1). Repeat the above steps 1), 2) and 3) in the control test.2). The first concentration of a test compound was pre-applied for 40sec followed by co-application of the test compound of the sameconcentration and EC₂₀ GABA to obtain I_(test). Divide I_(test) byI_(GABA) to obtain potentiation (%).3). Discard all tested oocytes, new oocytes were used and the abovesteps 1) and 2) were repeated to test second concentration of the samecompound. Each oocyte was used for only one concentration test for asingle test compound. The steps were repeated for other test compounds.

In some embodiments, the compounds of this application have a bindingaffinity (as represented by K_(i)) at α5-containing GABA_(A)Rs of lessthan 200 nM, less than 180 nM, less than 150 nM, or less than 100 nM. Insome embodiments, the compounds of this application have a bindingaffinity (as represented by K_(i)) at α5-containing GABA_(A)Rs of lessthan 50 nM. In some embodiments, the compounds of this application havea binding affinity (as represented by K_(i)) at α5-containing GABA_(A)Rsof less than 10 nM.

In some embodiments, the compounds of this application are selective forα5-containing GABA_(A)Rs over α1-containing GABA_(A)Rs. In someembodiments, the compounds of this application are more than 50-fold,more than 100-fold, more than 500-fold or more than 1000-fold selectivefor α5-containing GABA_(A)Rs over α1-containing GABA_(A)Rs.

In some embodiments, the compounds of this application have an EC₅₀ atthe α5-containing GABA_(A)Rs of less than 500 nM, less than 100 nM orless than 50 nM. In some embodiments, the compounds of this applicationhave an EC₅₀ at the α5-containing GABA_(A)Rs of less than 25 nM.

In some embodiments, the compounds of this application potentiateα5-containing GABA_(A)Rs for more than 10%, more than 25%, more than50%, or more than 75% at 100 nM. In some embodiments, the compounds ofthis application potentiate α5-containing GABA_(A)Rs for more than 10%,more than 25%, more than 50%, or more than 75% at 1000 nM.

Screening results of the binding and PAM functional activity tests aresummarized in Tables 1 and 2 below.

The following Table 1 illustrates the ranges of GABA α5 binding K_(i)'sassociated with compounds of this invention:

TABLE 1 GABA α5 Binding Ki Values (nM) <99 nM 100-1000 nM >1000 nMCompounds 1, 2, 3, Compounds 50, Compounds 116, 4, 6, 7,8, 9, 10, 11,110, 113, 115, 119, 117, 121, 123, 131, 12, 44, 55, 101, 103, 124, 125,134, 136, 135, 140, 142, 143, 105, 107, 108, 114, 138, 139, 141, 143,152, 154, 192, 193, 128, 153, 158, 162, 144, 146, 170, 191, 204, 221,229, 231, 163, 164, 166, 169, 200, 201, 219, 220, 234, 239, 250-253,171, 172, 173, 174, 237, 240, 246, 247, 262, 272, 279 175, 177, 179, 5,47, 248, 265-267, 273, 48, 49, 51, 52, 53, 274, 281, 283, 284, 54, 56,102, 104, 286, 287, 292, 297- 106, 111, 112, 118, 300, 303 120, 126,127, 130, 133, 137, 145, 147, 148, 149, 155, 156, 157, 160, 165, 168,178, 45, 46, 109, 122, 129, 132, 150, 151, 159, 161, 167, 176, 180-190,194-199, 202, 203, 205-210, 216, 217, 218, 222, 223-227, 230, 232, 233,235, 236, 238, 241-245, 249, 254-261, 263, 264, 268-271, 275- 278, 280,282, 285, 288-291, 293-296, 301, 302, 304

The following Table 2 illustrates the ranges of GABA α5 functionalpotentiation associated with compounds of this invention:

TABLE 2 GABA α5 Functional Data 20-49% @ 100 nM >50% @ 100 nM Compounds1, 2, 9, 11, Compounds 113, 48, 45, 55, 109, 110, 111, 114, 145, 149,160, 118, 120, 126, 127, 128, 171, 172, 173, 174, 130, 132, 137, 147,148, 176, 177, 178, 179, 153, 155, 158, 162, 163, 185, 186, 194, 271175, 180-184, 187-189, 191, 195, 196, 198, 199, 202, 203, 205, 207, 210,212, 213, 222, 224, 225, 226, 238, 243, 249, 254- 257, 264, 290, 293

Selected compounds of this invention demonstrate >10-fold bindingselectivity versus GABA α1, GABA α2, or GABA α3.

Example 107: Effect of Methyl 3,5-Diphenylpyridazine-4-Carboxylate inAged-Impaired (AI) Rats

Methyl 3,5-diphenylpyridazine-4-carboxylate, corresponding to compoundnumber 6 in van Niel et al. J. Med. Chem. 48:6004-6011 (2005), is aselective α5-containing GABA_(A) R agonist. It has an α5 in vitroefficacy of +27 (EC₂₀). The effect of methyl3,5-diphenylpyridazine-4-carboxylate in aged-impaired rats was studiedusing a RAM task. Moreover, receptor occupancy by methyl3,5-diphenylpyridazine-4-carboxylate in α5-containing GABA_(A) receptorwas also studied.

(A) Effect of Methyl 3,5-diphenylpyridazine-4-carboxylate inAged-Impaired Rats Using a Radial Arm Maze (RAM) Behavioral Task

The effects of methyl 3,5-diphenylpyridazine-4-carboxylate on the invivo spatial memory retention of aged-impaired (AI) rats were assessedin a Radial Arm Maze (RAM) behavioral task using vehicle control andfour different dosage levels of methyl3,5-diphenylpyridazine-4-carboxylate (0.1 mg/kg, 0.3 mg/kg, 1 mg/kg and3 mg/kg, ip). RAM behavioral tasks were performed on eight AI rats. Allfive treatment conditions (vehicle and four dosage levels) were testedon all eight rats.

The RAM apparatus used consisted of eight equidistantly-spaced arms. Anelevated maze arm (7 cm width×75 cm length) projected from each facet ofan octagonal center platform (30 cm diameter, 51.5 cm height). Clearside walls on the arms were 10 cm high and were angled at 650 to form atrough. A food well (4 cm diameter, 2 cm deep) was located at the distalend of each arm. Froot Loops™ (Kellogg Company) were used as rewards.Blocks constructed of Plexiglas™ (30 cm height×12 cm width) could bepositioned to prevent entry to any arm. Numerous extra maze cuessurrounding the apparatus were also provided.

The AI rats were initially subjected to a pre-training test (Chappell etal. Neuropharmacology 37: 481-487, 1998). The pre-training testconsisted of a habituation phase (4 days), a training phase on thestandard win-shift task (18 days) and another training phase (14 days)in which a brief delay was imposed between presentation of a subset ofarms designated by the experimenter (e.g., 5 arms available and 3 armsblocked) and completion of the eight-arm win-shift task (i.e., with alleight arms available).

In the habituation phase, rats were familiarized to the maze for an8-minute session on four consecutive days. In each of these sessions,food rewards were scattered on the RAM, initially on the center platformand arms and then progressively confined to the arms. After thishabituation phase, a standard training protocol was used, in which afood pellet was located at the end of each arm. Rats received one trialeach day for 18 days. Each daily trial terminated when all eight foodpellets had been obtained or when either 16 choices were made or 15minutes had elapsed. After completion of this training phase, a secondtraining phase was carried out in which the memory demand was increasedby imposing a brief delay during the trial. At the beginning of eachtrial, three arms of the eight-arm maze were blocked. Rats were allowedto obtain food on the five arms to which access was permitted duringthis initial “information phase” of the trial. Rats were then removedfrom the maze for 60 seconds, during which time the barriers on the mazewere removed, thus allowing access to all eight arms. Rats were thenplaced back onto the center platform and allowed to obtain the remainingfood rewards during this “retention test” phase of the trial. Theidentity and configuration of the blocked arms varied across trials.

The number of “errors” the AI rats made during the retention test phasewas tracked. An error occurred in the trial if the rats entered an armfrom which food had already been retrieved in the pre-delay component ofthe trial, or if the rat re-visited an arm in the post-delay sessionthat it had already visited.

After completion of the pre-training test, rats were subjected to trialswith more extended delay intervals, i.e., a two-hour delay, between theinformation phase (presentation with some blocked arms) and theretention test (presentation of all arms). During the delay interval,rats remained off to the side of the maze in the testing room, on cartsin their individual home cages. AI rats were pretreated 30-40 minutesbefore daily trials with a one-time shot of the following fiveconditions: 1) vehicle control—5% dimethyl sulfoxide, 25% polyethyleneglycol 300 and 70% distilled water; 2) methyl3,5-diphenylpyridazine-4-carboxylate at 0.1 mg/kg; 3) methyl3,5-diphenylpyridazine-4-carboxylate at 0.3 mg/kg; 4) methyl3,5-diphenylpyridazine-4-carboxylate at 1 mg/kg); and 5) methyl3,5-diphenylpyridazine-4-carboxylate at 3 mg/kg; through intraperitoneal(i.p.) injection. Injections were given every other day with interveningwashout days. Each AI rat was treated with all five conditions withinthe testing period. To counterbalance any potential bias, drug effectwas assessed using ascending-descending dose series, i.e., the doseseries was given first in an ascending order and then repeated in adescending order. Therefore, each dose had two determinations.

Parametric statistics (paired t-tests) was used to compare the retentiontest performance of the AI rats in the two-hour delay version of the RAMtask in the context of different doses of methyl3,5-diphenylpyridazine-4-carboxylate and vehicle control (see FIG. 1).The average numbers of errors that occurred in the trials weresignificantly fewer with methyl 3,5-diphenylpyridazine-4-carboxylatetreatment of 3 mg/kg (average no. of errors±standard error of the mean(SEM)=1.31±0.40) than using vehicle control (average no. oferrors±SEM=3.13±0.62). Relative to vehicle control treatment, methyl3,5-diphenylpyridazine-4-carboxylate significantly improved memoryperformance at 3 mg/kg (t(7)=4.233, p=0.004).

The therapeutic dose of 3 mg/kg became ineffective when the AI rats wereconcurrently treated with 0.3 mg/kg of TB21007, a α5-containing GABA_(A)R inverse agonist. The average numbers of errors made by rats with thecombined TB21007/methyl 3,5-diphenylpyridazine-4-carboxylate treatment(0.3 mg/kg TB21007 with 3 mg/kg methyl3,5-diphenylpyridazine-4-carboxylate) was 2.88±1.32, and was nodifferent from rats treated with vehicle control (3.13±1.17 averageerrors). Thus, the effect of methyl 3,5-diphenylpyridazine-4-carboxylateon spatial memory is a GABA_(A) α5 receptor-dependent effect (see FIG.1).

(B) Effect of Methyl 3,5-Diphenylpyridazine-4-Carboxylate onα5-Containing GABA_(A) Receptor Occupancy Animals

Adult male Long Evans rats (265-295 g, Charles River, Portage, Mich.,n=4/group) were used for GABA_(A)α5 receptor occupancy studies. Ratswere individually housed in ventilated stainless-steel racks on a 12:12light/dark cycle. Food and water were available ad libitum. Inadditional studies to evaluate compound exposures at behaviorally activedoses, young or aged Long Evan rats (n=2-4/group) were used for thesestudies.

Compounds

Ro 15-4513 was used as a receptor occupancy (RO) tracer for GABA_(A)α5receptor sites in the hippocampus and cerebellum. Ro 15-4513 was chosenas the tracer based on its selectivity for GABA_(A)α5 receptors relativeto other alpha subunit containing GABA_(A) receptors and because it hasbeen successfully used for GABA_(A)α5 RO studies in animals and humans(see, e.g., Lingford-Hughes et al., J. Cereb. Blood Flow Metab.22:878-89 (2002); Pym et al, Br. J. Pharmacol. 146: 817-825 (2005); andMaeda et al., Synapse 47: 200-208 (2003)). Ro 15-4513 (1 μg/kg), wasdissolved in 25% hydroxyl-propyl beta-cyclodextrin and administered i.v.20′ prior to the RO evaluations. Methyl3,5-diphenylpyridazine-4-carboxylate (0.1-10 mg/kg) was synthesized byNox Pharmaceuticals (India) and was dissolved in 25% hydroxyl-propylbeta-cyclodextrin and administered i.v. 15′ prior to tracer injection.Compounds were administered in a volume of 0.5 ml/kg except for thehighest dose of methyl 3,5-diphenylpyridazine-4-carboxylate (10 mg/kg)which was administered in a volume of 1 ml/kg due to solubilitylimitations.

Tissue Preparation and Analysis

The rats were sacrificed by cervical dislocation 20′ post tracerinjection. The whole brain was rapidly removed, and lightly rinsed withsterile water. Trunk blood was collected in EDTA coated eppendorf tubesand stored on wet ice until study completion. Hippocampus and cerebellumwere dissected and stored in 1.5 ml eppendorf tubes, and placed on wetice until tissue extraction. In a drug naïve rat, six cortical braintissues samples were collected for use in generating blank and standardcurve samples.

Acetonitrile containing 0.1% formic acid was added to each sample at avolume of four times the weight of the tissue sample. For the standardcurve (0.1-30 ng/g) samples, a calculated volume of standard reduced thevolume of acetonitrile. The sample was homogenized (FastPrep-24, LysingMatrix D; 5.5 m/s, for 60 seconds or 7-8 watts power using sonic probedismembrator; Fisher Scientific) and centrifuged for 16-minutes at14,000 rpm. The (100 μl) supernatant solution was diluted by 300 μl ofsterile water (pH 6.5). This solution was then mixed thoroughly andanalyzed via LC/MS/MS for Ro 15-4513 (tracer) and methyl3,5-diphenylpyridazine-4-carboxylate.

For plasma exposures, blood samples were centrifuged at 14000 rpm for 16minutes. After centrifuging, 50 ul of supernatant (plasma) from eachsample was added to 200 μl of acetonitrile plus 0.1% formic acid. Forstandard curve (1-1000 ng/ml) samples, a calculated volume of standardreduced the volume of acetonitrile. Samples were sonicated for 5 minutesin an ultrasonic water bath, followed by centrifugation for 30 minutes,at 16000 RPM. 100 ul of supernatant was removed from each sample vialand placed in a new glass auto sample vial, followed by the addition of300 μl of sterile water (pH 6.5). This solution was then mixedthoroughly and analyzed via LC/MS/MS for methyl3,5-diphenylpyridazine-4-carboxylate.

Receptor occupancy was determined by the ratio method which comparedoccupancy in the hippocampus (a region of high GABA_(A)α5 receptordensity) with occupancy in the cerebellum (a region with low GABA_(A)α5receptor density) and additionally by a high dose of the GABA_(A)α5negative allosteric modulator L-655,708 (10 mg/kg, i.v.) to define fulloccupancy.

Vehicle administration followed by tracer administration of 1 μg/kg,i.v., of Ro 15-4513 resulted in >5-fold higher levels of Ro 15-4513 inhippocampus (1.93±0.05 ng/g) compared with cerebellum (0.36±0.02 ng/g).Methyl 3,5-diphenylpyridazine-4-carboxylate (0.01-10 mg/kg, i.v.)dose-dependently reduced Ro 15-4513 binding in hippocampus, withoutaffecting cerebellum levels of Ro 15-4513 (FIG. 2) with a dose of 10mg/kg, i.v., demonstrating >90% occupancy (FIG. 3). Both methods ofcalculating RO yielding very similar results with ED50 values for methyl3,5-diphenylpyridazine-4-carboxylate as 1.8 mg/kg or 1.1 mg/kg based onthe ratio method or using L-755,608 to define occupancy.

Methyl 3,5-diphenylpyridazine-4-carboxylate exposure was below thequantification limits (BQL) at 0.01 mg/kg, i.v., in both plasma andhippocampus and but was detectable at low levels in hippocampus at 0.1mg/kg, i.v. (see Table 3). Hippocampal exposure was linear as a 10-foldincrease in dose from 0.1 to 1 mg/kg, i.v., resulted in a 12-foldincrease in exposure. Increasing the dose from 1 to 10 mg/kg, i.v., onlyincreased the exposure by ˜5-fold. Plasma exposure increased 12-fold asthe dose increased from 1 to 10 mg/kg, i.v.

TABLE 3 % GABA_(A) α5 Receptor Occupancy by methyl 3,5-diphenylpyridazine-4-carboxylate (0.01-10 mg/kg, i.v.). Hippocampus andPlasma Exposure of methyl 3,5- diphenylpyridazine-4-carboxylate byTreatment Group in young Long Evans rats. % RO % RO (L-655,708 (RatioPlasma Dose Method) Method) ng/mL Hippocampus (mg/kg, i.v.) (SEM) (SEM)(SEM) ng/g (SEM) 0.01 19.2 (11.1) 15.7 (9.1) BQL BQL 0.1 16.4 (4.9) 13.4 (4.0) BQL 14.6 (3.5) 1 38.5 (11.2) 31.5 (9.1) 62.8 (6.1) 180.0(10.3) 10 110.0 (6.6)   90.2 (5.4) 763.5 (85.7) 947.2 (51.3)

Additional studies were conducted in aged Long-Evans rats in order todetermine the exposures at the behaviorally relevant doses in thecognition studies. Exposure in young Long-Evans rats was also determinedto bridge with the receptor occupancy studies that were conducted inyoung Long-Evans rats. Exposures in young and aged Long-Evans rats wererelatively similar (Table 4, FIG. 4). Increasing the dose 3-fold from 1to 3 mg/kg, ip resulted in a greater than dose-proportional increase inexposure in young and aged rats in both hippocampus and plasma withincreases ranging from 4.5 to 6.6-fold.

TABLE 4 Hippocampus and Plasma Exposure of methyl3,5-diphenylpyridazine- 4-carboxylate in Young Long Evans Rats byTreatment Group Young Young Aged Aged Dose Hippocampus PlasmaHippocampus Plasma (mg/kg, ip) ng/g (SEM) ng/mL (SEM) ng/g (SEM) ng/mL(SEM) 1 25.9 (1.7) 20.0 (1.4)  38.8 (21.7) 45.2 (29.6) 3 129.1 (22.4)132.9 (19.5) 177.5 (19.5)  196 (18.2)

In the RO studies, an exposure of 180 ng/g in hippocampus (1 mg/kg,i.v.) represented 32-39% receptor occupancy depending on method used todetermine RO. This exposure is comparable to that observed in aged ratsat 3 mg/kg, i.p., suggesting that 30-40% RO is required for cognitiveefficacy in this model.

These studies demonstrated that methyl3,5-diphenylpyridazine-4-carboxylate produced dose-dependent increase inGABA_(A) α5 receptor occupancy. Methyl3,5-diphenylpyridazine-4-carboxylate also demonstrated good brainexposure with brain/plasma ratios >1. The studies further demonstratedthat methyl 3,5-diphenylpyridazine-4-carboxylate was producing itscognitive enhancing effects by positive allosteric modulation at theGABA_(A) α5 subtype receptor.

Example 108: Effect of Ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylatein Aged-Impaired (AI) Rats

Ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylate,corresponding to compound number 49 in Achermann et al. Bioorg. Med.Chem. Lett., 19:5746-5752 (2009), is a selective α5-containing GABA_(A)R agonist.

The effect of ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylateon the in vivo spatial memory retention of aged-impaired (AI) rats wasassessed in a Radial Arm Maze (RAM) behavioral task that is essentiallysimilar to the task as described in Example 107 (A), using vehiclecontrol (25% cyclodextrin, which was tested 3 times: at the beginning,middle and end of ascending/descending series) and six different doseslevels (0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 10 mg/kg and 30 mg/kg,each dose was tested twice) of ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylate.The same experiment was repeated using the same vehicle control anddoses of ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylate,where the vehicle control was tested 5 times, the 3 mg/kg dose of ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylatewas tested 4 times, and the other doses of ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylatewere tested twice.

Parametric statistics (paired t-tests) was used to compare the retentiontest performance of the AI rats in the four-hour delay version of theRAM task in the context of different doses of ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylateand vehicle control (see FIG. 5). Relative to vehicle control treatment,ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylatesignificantly improved memory performance at 3 mg/kg (t(7)=4.13,p=0.004, or t(7)=3.08, p=0.018) and at 10 mg/kg (t(7)=2.82, p=0.026).

The effect of ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylateon α5-containing GABA_(A) receptor occupancy was also studied followinga procedure that is essentially similar to the one as described inExample 107(B) (see above). This study demonstrated that ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylate(0.01-10 mg/kg, i.v.) reduced Ro 15-4513 binding in hippocampus, withoutaffecting cerebellum levels of Ro 15-4513 (FIG. 6) with a dose of 10mg/kg, i.v., demonstrating >90% occupancy (FIG. 7).

Example 109: Effect of 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one in Aged-Impaired Rats Using a Morris Water Maze Behavioral Task

6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one, corresponding to compound 44 in Chambers et al. J. Med. Chem.46:2227-2240 (2003) is a selective α5-containing GABA_(A) R agonist.

The effects of 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one on the in vivo spatial memory retention of aged-impaired (AI)rats were assessed in a Morris water maze behavioral task. A water mazeis a pool surrounded with a novel set of patterns relative to the maze.The training protocol for the water maze may be based on a modifiedwater maze task that has been shown to be hippocampal-dependent (de Hozet al., Eur. J. Neurosci., 22:745-54, 2005; Steele and Morris,Hippocampus 9:118-36, 1999).

Cognitively impaired aged rats were implanted unilaterally with acannula into the lateral ventricle. Stereotaxic coordinates were 1.0 mmposterior to bregma, 1.5 mm lateral to midline, and 3.5 mm ventral tothe skull surface. After about a week of recovery, the rats werepre-trained in a water maze for 2 days (6 trials per day) to locate asubmerged escape platform hidden underneath the surface of the pool, inwhich the escape platform location varied from day to day. Nointracerebroventricular (ICV) infusion was given during pre-training.

After pre-training, rats received ICV infusion of either 100 μg 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one (n=6) in μl DMSO or vehicle DMSO (n=5) 40 min prior to watermaze training and testing. Training consisted of 8 trials per day for 2days where the hidden escape platform remained in the same location.Rats were given 60 seconds to locate the platform with a 60 secondsinter-trial interval. The rats were given a probe test (120 seconds) 24hr. after the end of training where the escape platform was removed.During the training, there were 4 blocks, where each block had 4training trials.

Rats treated with vehicle and 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one found the escape platform about the same time at the beginningof training (block 1). In this block of training, rats treated withvehicle and 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one both spent about 24 seconds to find the escape platform.However, rats treated with 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one were able to find the platform more proficiently (i.e.,quicker) at the end of training (block 4) than those treated withvehicle alone. In block 4, rats treated with 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one spent about 9.6 seconds to find the escape platform, while ratstreated with vehicle spent about 19.69 seconds. These results suggestthat 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one improved the learning of the water maze task in rats (see FIG.8(A)).

During a test trial 24 hr. after training, the escape platform wasremoved. The search/swim pattern of the rats was used to measure whetherthe rats remember where the escape platform was located during pre-trialtraining in order to test for the long-term memory of the rats. In thistrial, “target annulus” is a designated area 1.5 times the size of theescape platform around the area where the platform was located duringpre-trial training. “Opposite annulus” is a control area of the samesize as the size of the target annulus, which is located opposite to thetarget annulus in the pool. If the rats had good long term memory, theywould tend to search in the area surrounding the location where theplatform was during the pre-trial training (i.e., the “target” annulus;and not the “opposite” annulus). “Time in annulus” is the amount of timein seconds that the rat spent in the target or opposite annulus area.“Number (#) of crossings” in annulus is the number of times the rat swamacross the target or opposite annulus area.

Rats received vehicle spent the same amount of time in the targetannulus and opposite annulus, indicating that these rats did not seem toremember where the platform was during the pre-trial training. Bycontrast, rats treated with 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one spent significantly more time in the target annulus, andcrossed the “target annulus” more often, as compared to the time theyspent in, or the number of times they crossed the “opposite annulus”.These results suggest that 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one improved the long-term memory of rats in the water maze task(see, FIGS. 8(B) and 8(C)).

Compounds of the present invention demonstrated positive allostericmodulatory effect on the GABA_(A) α5 receptor (See, e.g., Example 106).These compounds will enhance the effects of GABA at the GABA_(A) α5receptor. Therefore, compounds of the present invention should producecognitive enhancing effects in aged-impaired animals (such as rats),similar to the effects produced by other GABA_(A) α5 receptor selectiveagonists, such as methyl 3,5-diphenylpyridazine-4-carboxylate, ethyl3-methoxy-7-methyl-9H-benzo[f]imidazo[1,5-a][1,2,4]triazolo[4,3-d][1,4]diazepine-10-carboxylate,and 6,6dimethyl-3-(3-hydroxypropyl)thio-1-(thiazol-2-yl)-6,7-dihydro-2-benzothiophen-4(5H)-one (See, e.g., Examples 28-30).

1-11. (canceled)
 12. A method of treating cognitive impairment associated with a central nervous system (CNS) disorder in a subject in need thereof, comprising the step of administering a compound of Formula IV:

or a pharmaceutically acceptable salt, tautomer, stereoisomer, Z (zusammen) isomer, E (entgegen) isomer or combination thereof, wherein: R² is —OR⁸, —SR⁸, —(CH₂)_(n)OR⁸, —(CH₂)_(n)O(CH₂)_(n)R⁸, —(CH₂)_(p)R⁸ and —(CH₂)_(n)N(R″)R¹⁰; and wherein R² is independently substituted with 0-5 R′; m and n are independently integers selected from 0-4; p is an integer selected from 2-4; each occurrence of R¹, R⁴, and R⁵ are each independently selected from: halogen, —R, —OR, —NO₂, —NCS, —CN, —CF₃, —OCF₃, —SiR₃, —N(R)₂, —SR, —SOR, —SO₂R, —SO₂N(R)₂, —SO₃R, —(CR₂)₁₋₃R, —(CR₂)₁₋₃—OR, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃R, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃OR, —C(O)R, —C(O)C(O)R, —C(O)CH₂C(O)R, —C(S)R, —C(S)OR, —C(O)OR, —C(O)C(O)OR, —C(O)C(O)N(R)₂, —OC(O)R, —C(O)N(R)₂, —OC(O)N(R)₂, —C(S)N(R)₂, —(CR₂)₀₋₃NHC(O)R, —N(R)N(R)COR, —N(R)N(R)C(O)OR, —N(R)N(R)CON(R)₂, —N(R)SO₂R, —N(R)SO₂N(R)₂, —N(R)C(O)OR, —N(R)C(O)R, —N(R)C(S)R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂, —N(COR)COR, —N(OR)R, —C(═NH)N(R)₂, —C(O)N(OR)R, —C(═NOR)R, —OP(O)(OR)₂, —P(O)(R)₂, —P(O)(OR)₂, and —P(O)(H)(OR); R³ is absent or is selected from: halogen, —R, —OR, —NO₂, —NCS, —CN, —CF₃, —OCF₃, —SiR₃, —N(R)₂, —SR, —SOR, —SO₂R, —SO₂N(R)₂, —SO₃R, —(CR₂)₁₋₃R, —(CR₂)₁₋₃—OR, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃R, —(CR₂)₀₋₃—C(O)NR(CR₂)₀₋₃OR, —C(O)R, —C(O)C(O)R, —C(O)CH₂C(O)R, —C(S)R, —C(S)OR, —C(O)OR, —C(O)C(O)OR, —C(O)C(O)N(R)₂, —OC(O)R, —C(O)N(R)₂, —OC(O)N(R)₂, —C(S)N(R)₂, —(CR₂)₀₋₃NHC(O)R, —N(R)N(R)COR, —N(R)N(R)C(O)OR, —N(R)N(R)CON(R)₂, —N(R)SO₂R, —N(R)SO₂N(R)₂, —N(R)C(O)OR, —N(R)C(O)R, —N(R)C(S)R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂, —N(COR)COR, —N(OR)R, —C(═NH)N(R)₂, —C(O)N(OR)R, —C(═NOR)R, —OP(O)(OR)₂, —P(O)(R)₂, —P(O)(OR)₂, and —P(O)(H)(OR); each R⁶ is independently —H or —(C1-C6)alkyl; each R⁷ is independently —H or —(C1-C6)alkyl; each R⁸ is independently —(C1-C6)alkyl, —(C3-C10)-cycloalkyl, (C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each occurrence of R⁸ is independently substituted with 0-5 R′; each R¹⁰ is independently —(C3-C10)-cycloalkyl, 3- to 10-membered heterocyclyl-, (C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each occurrence of R¹⁰ is independently substituted with 0-5 R′; each R is independently selected from: H—, (C1-C12)-aliphatic-, (C3-C10)-cycloalkyl-, (C3-C10)-cycloalkenyl-, [(C3-C10)-cycloalkyl]-(C1-C12)-aliphatic-, [(C3-C10)-cycloalkenyl]-(C1-C12)-aliphatic-, [(C3-C10)-cycloalkyl]-O—(C1-C12)-aliphatic-, [(C3-C10)-cycloalkenyl]-O—(C1-C12)-aliphatic-, (C6-C10)-aryl-, (C6-C10)-aryl-(C1-C12)aliphatic-, (C6-C10)-aryl-O—(C1-C12)aliphatic-, (C6-C10)-aryl-N(R″)—(C1-C12)aliphatic-, 3- to 10-membered heterocyclyl-, (3- to 10-membered heterocyclyl)-(C1-C12)aliphatic-, (3- to 10-membered heterocyclyl)-O—(C1-C12)aliphatic-, (3- to 10-membered heterocyclyl)-N(R″)—(C1-C12)aliphatic-, 5- to 10-membered heteroaryl-, (5- to 10-membered heteroaryl)-O—(C1-C12)-aliphatic- and (5- to 10-membered heteroaryl)-N(R″)—(C1-C12)-aliphatic-; wherein said heterocyclyl has 1-4 heteroatoms independently selected from N, NH, O, S, SO, and SO₂, and said heteroaryl has 1-4 heteroatoms independently selected from N, NH, O, and S; wherein each occurrence of R is independently substituted with 0-5 R′; or when two R groups bound to the same atom, the two R groups may be taken together with the atom to which they are bound to form a 3- to 10-membered aromatic or non-aromatic ring having 0-4 heteroatoms independently selected from N, NH, O, S, SO, and SO₂, wherein said ring is optionally substituted with 0-5 R′, and wherein said ring is optionally fused to a (C6-C10)aryl, 5- to 10-membered heteroaryl, (C3-C10)cycloalkyl, or a 3- to 10-membered heterocyclyl; wherein each occurrence of R′ is independently selected from halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″, —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂; wherein each occurrence of R″ is independently selected from H, —(C1-C6)-alkyl, —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl, (C6-C10)-aryl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to 10-membered heteroaryl)-O—(C1-C6)-alkyl-, and (C6-C10)-aryl-O—(C1-C6)-alkyl-, wherein each occurrence of R″ is independently substituted with 0-3 substituents selected from: halogen, —R^(∘), —OR^(∘), oxo, —CH₂OR^(∘), —CH₂N(R^(∘))₂, —C(O)N(R^(∘))₂, —C(O)OR^(∘), —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R^(∘))₂, wherein each occurrence of R^(∘) is independently selected from: —(C1-C6)-aliphatic, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl, 5- to 10-membered heteroaryl-, and (C6-C10)-aryl-; or a pharmaceutical composition comprising a compound according to Formula (I), or a pharmaceutically acceptable salt, hydrate, solvate, polymorph, isomer, or combination thereof, in a therapeutically effective amount; and an acceptable carrier, adjuvant or vehicle. 13-58. (canceled)
 59. A method of treating cognitive impairment associated with a central nervous system (CNS) disorder in a subject in need thereof, comprising the step of administering a compound or a pharmaceutical composition according to claim 12, or a pharmaceutically acceptable salt, tautomer, stereoisomer, Z (zusammen), E (entgegen) isomer, or combination thereof, wherein: m is 0-3; each R¹ is independently selected from: halogen, —H, —(C1-C6)alkyl, —C≡CH, —OH, —O((C1-C6)alkyl), —NO₂, —CN, —CF₃, —OCF₃, wherein R¹ is independently substituted with 0-5 R′; R² is selected from —OR⁸, —SR⁸, —(CH₂)_(n)OR⁸, —(CH₂)_(n)O(CH₂)_(n)R⁸, —(CH₂)_(p)R⁸ and —(CH₂)_(n)N(R″)R¹⁰, wherein n is an integer selected from 0-4; p is an integer selected from 2-4; each R⁸ is independently —(C1-C6)alkyl, —(C3-C10)-cycloalkyl, (C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each occurrence of R⁸ is independently substituted with 0-5 R′; each R¹⁰ is independently —(C3-C10)-cycloalkyl, 3- to 10-membered heterocyclyl-, (C6-C10)-aryl, or 5- to 10-membered heteroaryl, wherein each occurrence of R¹⁰ is independently substituted with 0-5 R′; and wherein R² is independently substituted with 0-5 R′; R³ is selected from: H, —CN, halogen (e.g., Br), —(C1-C6)alkyl, —C≡CH, —SO₂((C1-C6)alkyl), —C(O)N((C1-C6)alkyl)₂), —C(O)NH((C1-C6)aliphatic)₂, (C6-C10)-aryl-(C1-C12)aliphatic-, —C(O)((C1-C6)alkyl), —C(O)O((C1-C6)alkyl), 5- or 6-membered heterocyclyl, and 5- or 6-membered heteroaryl; and wherein R³ is independently substituted with 0-5 R′; R⁴ and R⁵ are each independently selected from —H, halogen and —(C1-C6)alkyl; R⁶ is selected from —H and —(C1-C6)alkyl; wherein each occurrence of R′ is independently selected from halogen, —R″, —OR″, oxo, —CH₂OR″, —CH₂NR″₂, —C(O)N(R″)₂, —C(O)OR″, —NO₂, —NCS, —CN, —CF₃, —OCF₃ and —N(R″)₂; wherein each occurrence of R″ is independently selected from H, —(C1-C6)-alkyl, (C3-C6)-cycloalkyl, 3- to 6-membered heterocyclyl, (C6-C10)-aryl-, (C6-C10)-aryl-(C1-C6)-alkyl-, (5- to 10-membered heteroaryl)-O—(C1-C6)-alkyl-, and (C6-C10)-aryl-O—(C1-C6)-alkyl-.
 60. The method according to claim 12, wherein the compound is selected from: Cmp No. Structure 215

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or a pharmaceutically acceptable salt, hydrate, solvate, polymorph, isomer, or combination thereof.
 61. The method according to claim 12, wherein the compound is selected from Compound Structure 183

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or a pharmaceutically acceptable salt, hydrate, solvate, polymorph, isomer, or combination thereof.
 62. The method according to claim 12, wherein the CNS disorder is age-related cognitive impairment.
 63. The method according to claim 12, wherein the CNS disorder is mild cognitive impairment (MCI).
 64. The method according to claim 12, wherein the CNS disorder is amnestic mild cognitive impairment (aMCI).
 65. The method according to claim 12, wherein the CNS disorder is dementia.
 66. The method according to claim 12, wherein the CNS disorder is Alzheimer's disease.
 67. The method according to claim 12, wherein the CNS disorder is selected from the group consisting of schizophrenia, bipolar disorder, amyotrophic lateral sclerosis (ALS), post-traumatic stress disorder (PTSD), mental retardation, Parkinson's disease (PD), autism, compulsive behavior, substance addiction.
 68. The method according to claim 60, wherein the CNS disorder is age-related cognitive impairment.
 69. The method according to claim 60, wherein the CNS disorder is mild cognitive impairment (MCI).
 70. The method according to claim 60, wherein the CNS disorder is amnestic mild cognitive impairment (aMCI).
 71. The method according to claim 60, wherein the CNS disorder is dementia.
 72. The method according to claim 60, wherein the CNS disorder is Alzheimer's disease.
 73. The method according to claim 60, wherein the CNS disorder is selected from the group consisting of schizophrenia, bipolar disorder, amyotrophic lateral sclerosis (ALS), post-traumatic stress disorder (PTSD), mental retardation, Parkinson's disease (PD), autism, compulsive behavior, substance addiction.
 74. The method according to claim 61, wherein the CNS disorder is age-related cognitive impairment.
 75. The method according to claim 61, wherein the CNS disorder is mild cognitive impairment (MCI).
 76. The method according to claim 61, wherein the CNS disorder is amnestic mild cognitive impairment (aMCI).
 77. The method according to claim 61, wherein the CNS disorder is dementia.
 78. The method according to claim 61, wherein the CNS disorder is Alzheimer's disease.
 79. The method according to claim 61, wherein the CNS disorder is selected from the group consisting of schizophrenia, bipolar disorder, amyotrophic lateral sclerosis (ALS), post-traumatic stress disorder (PTSD), mental retardation, Parkinson's disease (PD), autism, compulsive behavior, substance addiction. 